Diagnostic Value of Karyotyping, CMA/CNV-Seq, and WES in Fetuses with Thickened Nuchal Translucency: Perinatal and Two-Year Follow-Up Outcomes

Diagnostic Value of Karyotyping, CMA/CNV-Seq, and WES in Fetuses with Thickened Nuchal Translucency: Perinatal and Two-Year Follow-Up Outcomes | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Diagnostic Value of Karyotyping, CMA/CNV-Seq, and WES in Fetuses with Thickened Nuchal Translucency: Perinatal and Two-Year Follow-Up Outcomes Mohan Wang, Yizhen Ji, Yasong Xu, Shiyu Sun, Xiaomei Yang, Li Sun, and 1 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-8072036/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 21 Feb, 2026 Read the published version in BMC Medical Genomics → Version 1 posted 12 You are reading this latest preprint version Abstract Background This study aimed to analyze the perinatal and pediatric outcomes of fetuses with thickened nuchal translucency (NT ≥ 2.5 mm) to enhance prenatal diagnostic strategies. Study Design A total of 720 pregnant women with NT ≥ 2.5 mm in the first trimester underwent interventional prenatal diagnosis. These participants were followed up during the perinatal and pediatric periods (2 years after birth). Results The incidence of fetal chromosomal karyotype abnormalities was 32.86%, with trisomy 21 being the most common abnormality. Chromosomal microarray analysis (CMA) and copy number variation sequencing (CNV-seq) increased the detection rate by 10.72%. The most prevalent pathogenic copy number variations (pCNVs) and likely pathogenic copy number variations (lpCNVs) were associated with 22q11.21 microdeletion/duplication syndrome and 15q11.2 microdeletion syndrome, respectively. Excluding pathogenic karyotype abnormalities and pCNV/lpCNVs, the rate of pathological deformities was 19.6%. Whole-exome sequencing (WES) was performed in 11 cases, yielding a detection rate of 72.7%. There were 339 pregnancy terminations and 381 live births, of which 337 had normal karyotypes, CNVs, and ultrasound results, resulting in a live-birth incidence of 96.8%. In addition, two cases of psychomotor retardation were identified. Conclusion Traditional karyotyping, CMA/CNV-seq testing, and detailed ultrasound examinations are vital diagnostic tools that support genetic counseling for fetuses with thickened NT in the first trimester. Therefore, novel and efficient WES testing is required. Nuchal translucency Chromosome karyotype CMA/CNV-seq Perinatal outcomes Figures Figure 1 Key Points 1. Thickened NT linked to high chromosomal risk 2. CMA/CNV-seq improved detection beyond karyotype 3. WES showed high yield in selected cases Background Nuchal translucency (NT) refers to fluid accumulation behind the neck of a developing fetus, which can be measured via ultrasound during early pregnancy. It serves as an important indicator of potential chromosomal abnormalities, deformities, fetal anemia, and hypoproteinemia [ 1 – 3 ]. On ultrasound images, the NT appears as an echoless area behind the neck between the deep soft tissue hyperechoic band and skin hyperechoic zone [ 2 ]. The fundamental pathophysiological mechanisms underlying NT thickening are complex and not completely understood. The primary cause of NT thickening in fetuses is associated with cardiac anatomical abnormalities. Moreover, abnormalities in the lymphatic system and reflux disorders contribute to heart failure due to congenital malformations and genetic factors [ 3 ]. In 1992, Nicolaides et al. first proposed the use of fetal NT thickening in the first trimester as a marker of Down syndrome [ 4 ]. However, the International Society of Obstetrics and Gynecology Ultrasound currently lacks a strict definition for the range of NT thickening, and the specific cut-off value for abnormal NT ranges remains controversial in various prenatal diagnostic centers, both domestically and internationally [ 5 ]. Most clinical practices in China currently adopt NT ≥ 2.5 mm as the cutoff. Recent advances in research and the introduction of novel genetic testing technologies have revealed that NT thickening, even in the absence of chromosomal abnormalities, is associated with adverse pregnancy outcomes. These outcomes include fetal chromosomal microdeletion and microduplication syndromes (MMS), structural anomalies, certain genetic syndromes, intrauterine fetal death, and postnatal neuromotor developmental delays [ 6 ]. However, NT thickening is not directly correlated with fetal abnormalities. Research indicates that when fetal karyotype and microdeletion/microduplication tests yield normal results, the complete absence of deformities and growth retardation of the fetus in a systematic ultrasound examination suggests a high probability of a positive pregnancy outcome (> 95%) [ 7 ]. NT thickening and subsequent diagnostic examination are major concerns in pregnant women. The intricate association between NT and chromosomal abnormalities necessitates extensive prenatal genetic counseling and individualized diagnostic programs for pregnant women. Currently, informing pregnant women and their families about accurate fetal and pregnancy outcomes is crucial for improving their reassurance, whereas providing clinicians with reliable data to mitigate the residual risk of postpartum issues remains a major challenge in maternal-fetal medicine. However, there is no consensus on the approaches to prenatal diagnosis and management of these conditions. This study retrospectively analyzed 720 singleton cases of interventional prenatal diagnoses associated with an NT of ≥ 2.5 mm in the first trimester. The analysis included copy number variations (CNV), whole-exome sequencing (WES), and fetal karyotype assessments. Additionally, this study examined fetal ultrasound screenings for deformities, tracked pregnancy outcomes, and monitored the growth and development of live infants for up to two years after birth. This study aimed to provide guidance for clinical consultation, prenatal diagnosis strategies, prognostic evaluation, and reproductive planning in cases of increased fetal NT thickness. Materials and Methods Subjects This study retrospectively analyzed 720 singleton cases of interventional prenatal diagnosis. The cohort comprised 342 primiparous and 378 multiparous mothers who underwent prenatal examinations and ultrasonography between 11 and 13 + 6 weeks of pregnancy, revealing a fetal NT ≥ 2.5 mm. Data were collected from January 1, 2018, to December 31, 2021. The average age of pregnant women meeting the inclusion criteria was 31.08 (18–46 years). Pregnancies associated with identified chromosomal abnormalities or significant defects diagnosed prenatally or postnatally were excluded from the analysis. In total, 213 patients underwent transabdominal chorionic villus sampling, 483 underwent transabdominal amniocentesis, and 24 underwent transabdominal umbilical cord blood sampling. All interventional prenatal diagnoses were performed, and the samples were analyzed using G-banded chromosome karyotyping and CMA/CNV-seq detection technology. Among these cases, 11 with normal fetal karyotypes and CMA/CNV-seq results were further tested using WES due to deformities. For fetuses with identified karyotypes or copy number abnormalities, couples are recommended to provide peripheral blood samples to further classify the variants as de novo or genetic. All pregnant women and their families were informed of the indications and risks associated with interventional punctures before surgery and signed an "Informed Consent Form for Surgery.” Following the acquisition of informed consent, the study was approved by the Ethics Committee of Women and Children’s Hospital of Xiamen University. Fetal NT measurement standards Ultrasound measurement of NT thickness is based on the British Fetal Foundation Medical Society (FMF) standards for gestational ages between 11 and 13 + 6 weeks or when the fetal crown-rump length is approximately 45–84 mm [ 8 ]. In this study, an NT measurement of ≥ 2.5 mm was considered abnormal. High-resolution ultrasound was used, positioning the probe perpendicular to the skin of the fetal neck and back to ensure that the fetus was in a natural propensity position. A median sagittal section was obtained, and the long ribbon-like anechoic area beneath the skin of the fetal neck and back was measured using a Vernier caliper. The measurements were repeated thrice, and the maximum value was recorded as the NT thickness. CMA/CNV-seq detection In this study, CMA was conducted in collaboration with the Beijing Beikang Medical Laboratory, and CNV-seq was performed at the BGI Genomics Medical Laboratory. All participants signed an informed consent form after a comprehensive explanation of the CMA/CNV-seq detection technology. Samples included 5–10 mg of chorionic villi, 10 ml of amniotic fluid, and 2 ml of umbilical cord blood, which were sent for testing. Whole exome sequencing Briefly, 5–10 mg of chorionic villi, 10 ml of amniotic fluid, and 2 ml of umbilical cord blood were collected for testing. Genomic DNA was extracted using a commercial kit. Genomic DNA was fragmented, purified, and labeled to establish the library. A MGISEQ-2000 high-throughput sequencer was used for sequencing with a PE100 + 10 sequencing type, and the original sequencing data were obtained upon completion. Sequences with HG19/HG20 reference genomes were aligned using the Burrows-Wheeler Aligner (BWA) software. The sequence capture effect was concurrently evaluated, and single nucleotide variant (SNV) and insertions/deletions (Indels) were identified using the GATK software to generate the base polymorphism results for the target region. Suspicious mutations were annotated and screened, and Sanger sequencing was performed to verify all identified pathogenic mutations, confirming the microarray capture and high-throughput sequencing results. According to the latest version of the American College of Medical Genetics and Genomics (ACMGG) genetic variation classification standards and guidelines (2015), CNVs were categorized into five groups [ 9 ]: benign CNVs (bCNVs), likely benign CNVs (lbCNVs), variants of uncertain significance CNVs (VOUS CNVs), likely pathogenic CNVs (lpCNVs), and pathogenic CNVs (pCNVs). Results Chromosome karyotype test results In this study, 720 samples were subjected to G-banded chromosome karyotyping. Successful cultures were obtained from 715 cases, with 480 demonstrating normal karyotypes and 235 showing abnormal karyotypes, resulting in an abnormality rate of 32.86%. This included 211 cases with abnormal chromosome numbers: 161 cases with abnormal autosome numbers, with trisomy 21 being the most common, followed by trisomy 18 and trisomy 13. There were 50 cases of sex chromosome number abnormalities, with 45 X being the most prevalent. Additionally, 24 cases of structural chromosomal abnormalities were identified; 21 cases had unbalanced structural abnormalities, and three cases had balanced structural abnormalities (Table 1 ). Table 1 Distribution of karyotypes of 235 cases of NT thickened chromosomal abnormalities Karyotype type Number of cases Abnormal chromosome number 211 Abnormal number of autosomes 161 47, XN, + 21 106 47, XN, + 18 42 47, XN, + 13 12 47, XN, +7 1 Abnormal number of sex chromosomes 50 45, X 40 47, XXX 2 47, XXY 5 47, XYY 3 Chromosome structural abnormalities 24 Balance 46, XN, ?inv (18) (p11.3q11.2) 46, XN, t (1;9) (q42.1;q32) 46, XN, t (7;14) (q31;q13) mat, 3 Imbalance 46, XN, del (1) (q23;q24) 46, XN, del (14) (q24.2q31.2) 46, XN, del (18) (p11.2) dn 46, XN, del (4) (p16.1) 46, XN, del (10) (q26.1) dn 46, XN, del (4) (q13.1q22) 46, XN, del (15) (q11.2q13.1) 46, XN, del (11) (q13.5q23.1) dn 46, XN, del (4) (p15.1) dn 46, XN, del (2) (q36.3) [ 14 ]/46, XN [ 6 ] 46, XN, ? dup (1) (q32q41) dn 46, XN, der (10) t (10;22) 46, XN, der (8) t (5;8) (p12;p23.1) pat 46, XN, der (1) t (1;4) (p36.2;q31.2) pat 46, XN, der (11) t (6;11) (q23;q25) mat 46, XN, rec (18) dup (18q) inv (18) (p11.32q12.1) mat 46, XN, inv (9) (p11q13), dup (11) (q24q13) 47, XN, +? i (12) (p10) dn 46, XN, i (18) (q10) [34]/46, XN, del (18) (p11.1) [ 14 ] 46, X, psu dic (Y) (q11.22 [113]/45, X [ 10 ] 47, XN, +mar [33]/46, XN [ 15 ] dn 21 Note: 47, XN, + 21 contains 4 chimerism; 47, XN, + 18 contains 1 chimerism; 47, XN, + 13 contains 1 chimerism; 45, X contains 6 chimerism; 47, XXX contains 1 chimerism. CMA/CNV-seq detection results CMA/CNV-seq successfully detected abnormalities in 211 cases with chromosomal abnormalities and 21 cases with structural chromosomal imbalances, as identified by karyotype analysis. Additionally, CMA/CNV-seq identified pCNVs in five samples in which karyotype culture failed to identify them. Among 480 cases with normal karyotypes, CMA/CNV-seq detected 23 pCNVs and 24 lpCNVs, resulting in a 10.72% increase in the detection rate compared with karyotyping. 22q11.21 MMS was the most common (26.09%) of the 23 pCNVs, followed by Yq11.223q11.23 microdeletion syndrome (17.39%) (Table 2 ). Copy number variations associated with neurocognitive disorders were detected in all 24 lpCNV cases, with 15q11.2 microdeletion syndrome being the most common (37.5%), characterized by CNVs ranging from 311.8kb to 861.33kb. Other notable CNVs included 16p13.11 MMS (12.5%) with sizes from 1.63Mb ~ 3.29Mb, and 22q11.21 MMS (12.5%) ranging from 114.86kb ~ 1.58Mb. Two cases of Xp22.31 micro-repeats exhibited CNV sizes of 1.6 Mb (Table 3 ). Table 2 PCNVs detection in 23 fetuses with thickened NT and normal karyotype Number NT mm Ultrasound results CNV area and size CNV involves genes and syndromes/diseases 1 4.2 Congenital heart disease del1p36.33p36.22(10.0Mb) Contains 103 OMIM genes including GABRD/1p36 deletion syndrome 2 2.9 No obvious structural deformity was seen del3q29q29(1.69Mb)dn Contains 21 protein-coding genes including DLG1/3q29 deletion syndrome 3 3 No obvious structural deformity was seen del6p25.3p22.2(25.3Mb) del6q27(2.5Mb) Contains 89 OMIM genes including FOXC1/6pter-p24 deletion syndrome Contains 14 OMIM genes including ERMARD/related to AD hereditary periventricular nodular ectopia and other diseases 4 5 No obvious structural deformity was seen del7q11.23(1.5Mb)dn Contains 26 OMIM genes including ELN/related to Williams-Beuren syndrome (AD) 5 4.1 No obvious structural deformity was seen del10p15.3p15.1(4.8Mb)dn Contains 9 OMIM genes including ZMYND11/related to autosomal dominant intellectual disability 6 2.6 Congenital heart disease dup10p12.2p11.23(5.7Mb) Contains 22 OMIM genes including ANKRD26/related to diseases such as thrombocytopenia type 2 (AD) 7 3.2 No obvious structural deformity was seen del14q32.12q32.13(1.63Mb)pat Contains 22 protein-coding genes/associated with pleuropulmonary blastoma 8 2.7 Short limbs Uniparental diploid (UPD) on chromosome 14 Contains DLK1 and other genes/related to Temple syndrome (mat) and Kagami-Ogata syndrome (pat) 9 3.1 Bilateral polycystic kidney disease del17q12(1.4Mb) Contains 17 OMIM genes including HNF1B/recurrent 17q12 deletion syndrome 10 2.78 Left polycystic kidney, right hydronephrosis del17q12(1.5Mb)dn Contains 12 OMIM genes including HNF1B/recurrent 17q12 deletion syndrome 11 2.8 Bilateral polycystic kidney disease del17q12q12(1.99Mb)dn This region contains 26 protein-coding genes/17q12 deletion syndrome 12 8.09 congenital heart disease del22q11.21(3.1Mb) Contains 44 OMIM genes including TBX1/22q11.2 deletion syndrome 13 2.9 congenital heart disease del22q11.21(3.1Mb)dn Contains 44 OMIM genes including TBX1/22q11.2 deletion syndrome 14 4.7 No obvious structural deformity was seen del22q11.21(2.8Mb) Contains 42 OMIM genes including TBX1/22q11.2 deletion syndrome 15 2.9 No obvious structural deformity was seen del22q11.21q11.23(2.6Mb) Contains 30 OMIM genes including BCR/22q11.2 distal deletion syndrome 16 3.37 No obvious structural deformity was seen dup22q11.21(2.8Mb)dn Contains 41 OMIM genes including CLTCL1/22q11.2 microduplication syndrome 17 3.94 No obvious structural deformity was seen dup22q11.21q11.21(2.25Mb) This region contains 37 protein-coding genes/22q11.2 duplication syndrome 18 2.8 Intrauterine growth retardation dupXp22.31(1.6Mb) Contains 4 OMIM genes including STS/related to clinical phenotypes such as ichthyosis, global developmental delay, autism, and epilepsy 19 7.82 Neck hygroma, generalized edema delXp22.33q28(155.27Mb) Associated with Turner syndrome 20 5.7 Neck hygroma, generalized edema delYq11.223q11.23(2.25Mb) Contains 17 protein-coding genes including AZFc and DAZ regions/Y-linked spermatogenesis disorder 21 3.49 No obvious structural deformity was seen delYq11.223q11.23(3.11Mb) This region contains 13 protein-coding genes/Y-linked spermatogenesis disorders. 22 3 No obvious structural deformity was seen delYq11.223q11.23(2.16Mb) Contains 7 protein-coding genes including BPY2/Y-linked spermatogenesis disorder 23 3.1 No obvious structural deformity was seen delYq11.223q11.23(3.55Mb) pat Contains 10 protein-coding genes including BPY2/Y-linked spermatogenesis disorder Table 3 Detection results of LPCNVs in 7 fetuses with thickened NT and normal karyotype Number NTmm CNV area and size CNV involves genes and syndromes/diseases 1 6.4 dup4q22.1(1.8Mb) Contains 3 OMIM genes including SNCA/related to diseases such as dementia with Lewy bodies (AD) 2 3.2 del5p15.33p15.33(1.05Mb)dn 18 protein-coding genes, including the SDHA gene, are related to diseases such as delayed optic atrophy (AD) 3 4 dup5p15.1p14.1(8.6Mb) Contains 5 OMIM genes including CDH18/related to clinical phenotypes such as autism, global developmental delay, and epilepsy 4 4.3 dup6q25.3(2.4Mb) Contains 10 OMIM genes including ARID1B/related to CoffinSiris syndrome (AD) disease, and clinical phenotypes include intellectual disability, alopecia, sensorineural deafness, etc. 5 2.9 dup7p21.3(3.9Mb) Contains TMEM106B and other 5 OMIM genes/related to hypomyelination leukodystrophy 16 (AD) disease, clinical phenotypes include intellectual disability, learning disabilities, epileptic seizures, gait ataxia, etc. 6 3.24 del20p12.3(1.4Mb) Involving PLCB1 and other 3 OMIM genes/associated with early infantile epileptic encephalopathy type 12 (AR) 7 3.3 delXq28(376.0Kb) Containing part of the AFF2 gene segment/FRAXE type intellectual disability (X-linked recessive), clinical phenotypes include mild to moderate intellectual disability, cognitive difficulties, communication disorders, ADHD, autistic behavior, etc. Excluding the correlation between pathogenic chromosomal karyotypic abnormalities and pCNVs/lpCNVs, NT thickening and pathological deformities In this study, pathogenic chromosomal karyotype abnormalities and pCNV/lpCNVs were excluded, and 85 cases of NT-thickened fetal ultrasound examinations were found to have pathological deformities (19.6%). Among them, 24 presented with fetal neck cystic hygroma, generalized skin edema, and bilateral pleural effusion. The remaining 61 patients had deformities, including 17 with multiple malformations and 44 with single malformations. Systemic deformities were classified as follows: 25 cases of cardiac malformations, 19 cases of skeletal system deformities, and 18 cases of thoracic and abdominal deformities, including eight cases of omphalocele, seven cases of gastroschisis, one case of diaphragmatic hernia, one case of duodenal atresia, and one case of an abdominal mass. Additionally, eight cases of renal dysplasia and seven cases of abnormal nervous system development were identified. Cardiac malformations were the most frequently observed structural abnormality. Among the 11 fetuses with pathological deformities tested using WES, eight were found to have pathogenic or possibly pathogenic gene mutations, resulting in a detection rate of 72.7%. Two patients had similar abnormal ultrasound results on prior examinations. WES identified monogenic diseases consistent with autosomal recessive inheritance. Ultrasonography in three cases showed abnormal skeletal development, and pathogenic genes associated with skeletal dysplasia were detected. The remaining three cases exhibited pathogenic genes linked to abnormal phenotypes (Table 4 ). Table 4 The karyotype and CMA/CNV-seq of 8 fetal cases with thickened NT were normal and WES detected pathogenic/suspected pathogenic variants Number NT mm Ultrasound test results Adverse reproductive history Mutated genes/associated diseases/inheritance patterns 1 3.4 Short limb deformity, narrow chest and large abdomen None FGFR3:c.2420G > T(p.807Leufs*?) dn Lethal osteoplasia/AD 2 12.7 Generalized skin edema and bone dysplasia None COL2A1:c.3022G > C (p.Gly1008Arg)dn osteoarthritis with mild achondroplasia/AD 3 5.4 Neck hygroma, systemic skin edema, short limbs, abnormal intracardiac structure None SLC26A2:c.2018A > T(p.Asp673Val) heterozygous (inherited from father) SLC26A2:c.2046G > A(p.Leu682Leu) heterozygous (inherited from mother) Deformative bone dysplasia, chondrogenesis type 1B/AR 4 4.79 Omphalocele, ventricular septal defect Similar ultrasound abnormalities recur PIGN:c.963G > A(p.Gln321Gln) The fetus is homozygous (inherited from both father and mother) Multiple congenital malformations-hypotonia-epilepsy syndrome type 1/AR 5 7.35 Generalized skin edema Past unexplained neonatal death LZTR1:c.742G > A(p.Gly248Arg) (inherited from mother) Noonan syndrome/AD 6 6.9 Generalized skin edema Similar ultrasound abnormalities occur again CHRNA1:c.1128delG(p.Pro377Leufs*10) heterozygous (inherited from mother) CHRNA1:c.505T > C(p.Trp169Arg) heterozygous (inherited from father) Multiple fatal pterygium syndrome/AR 7 9 Generalized skin edema None KMT2D:c.9769A > T(p.Lys3257*)dnKabuki syndrome type 1/AD 8 6.98 Omphalocele, general skin edema None MAP2K2:c.1073_1074dupCG(p.Asp359Argfs*3)dncardiofacial-cutaneous syndrome type 4/AD Analysis of perinatal outcomes of 720 cases of fetal NT thickening Among the 720 singleton pregnancies, there were 339 pregnancy terminations and 381 live births (Fig. 1 ). In this study, 211 cases of chromosomal abnormalities and 20 cases of structural chromosomal imbalances diagnosed via karyotype analysis were terminated, accounting for 68.1% of terminations. Additionally, 71 cases without karyotype abnormalities or pCNV/lpCNVs but with pathological deformities detected via ultrasound accounted for 20.9% of the cases. Furthermore, 21 cases of pCNVs (including five cases in which karyotype culture failed) and five cases of lpCNVs detected by CMA/CNV-seq led to pregnancy termination (7.7%). Eleven pregnancies with normal fetal karyotypes, CNVs, and ultrasound results were terminated, accounting for 3.2% of all pregnancies. Among these, five cases were miscarriages, one was a stillbirth, three were diagnosed with severe α-thalassemia, and two cases of premature intrauterine growth restriction and abnormal umbilical blood flow required termination. Among the 381 live births, 337 had normal fetal karyotypes, CNVs, and ultrasound findings. Of these, 317 were full-term deliveries and 20 were premature births. The birth weight of these newborns was 3367.0 ± 450.6 g, resulting in a live birth rate of 96.8%. The remaining 44 live births included 14 cases of pathological deformities detected via ultrasound, excluding karyotype abnormalities and pCNVs/lpCNVs. There were also 19 cases of lpCNVs, three cases of balanced chromosomal translocations with no ultrasound abnormalities, seven cases of pCNVs, and one case of unbalanced chromosomal translocation. Analysis of the follow-up status of 381 live-born infants in early childhood All live births were followed up in early childhood within the first two years, during which a pediatrician performed clinical neurodevelopmental assessments. Five cases of psychomotor retardation were identified. One case involved del22q11.21 (2.8Mb), which exhibited no obvious deformities on prenatal ultrasound but was later diagnosed with congenital heart disease (atrial septal defect) after birth. By two years of age, the child exhibited psychomotor retardation and mental impairments. Another case involved dupXp22.31 (1.6Mb), with prenatal ultrasound showing fetal growth retardation and polyhydramnios. This infant was born full-term via vaginal delivery with a birth weight of 2050 g, and additional WES testing confirmed Noonan syndrome. In another case, prenatal ultrasound suggested a possible partial absence of the corpus callosum, which was later confirmed by ultrasound at six months post-birth, alongside a diagnosis of congenital heart disease (atrial septal defect). WES also suggested Noonan syndrome in this case. Among the 337 live births with normal karyotypes, CNVs, and ultrasound results, one case had an NT measurement of 8.69 mm in the first trimester. WES testing identified a heterozygous mutation (dn) in SETD5 associated with autosomal dominant intellectual developmental disorder-23 (MRD23). Another case, with an NT measurement of 9.1 mm in the first trimester, was found to have a SOS1 gene mutation (dn) associated with Noonan syndrome type 4. Discussion This study involved 720 pregnant women with first-trimester NT measurements ≥ 2.5 mm, each undergoing interventional prenatal diagnosis with follow-up during the perinatal period and early childhood (up to 2 years after birth). The results confirm that early fetal NT thickening is associated with adverse pregnancy outcomes, such as chromosomal abnormalities, chromosomal MMS, structural fetal anomalies, specific genetic syndromes, intrauterine fetal death, and postnatal neuromotor development delays. In this study, the rate of fetal karyotype abnormalities was as high as 32.86%, with trisomy 21 being the most common, highlighting a strong association between fetal NT thickening and chromosomal abnormalities, particularly in Down syndrome. A domestic study using a 2.5 mm NT threshold and amniotic fluid karyotype analysis of 4028 pregnant women reported a chromosomal abnormality rate of 22.02% among fetuses with NT thickening, which was significantly higher than that in fetuses without NT thickening, with trisomy 21 being the most prevalent [ 10 ]. Moreover, even in the absence of chromosomal abnormalities, fetuses with NT thickening face heightened risks of a wide range of congenital anomalies, including isolated structural defects, genetic syndromes, and neurodevelopmental delays that often manifest in early childhood [ 11 ]. A Meta-analysis revealed that CMA technology could detect an additional 5% of pathogenic variants in cases of fetal NT thickening, with 22q11.2 MMS, 10q26.12q26.3 deletions, and 12q21q22 deletions identified as the most frequent pathogenic CNVs [ 12 ]. Our results confirmed that after excluding chromosomal karyotype abnormalities, fetal NT thickening was associated with MMS in the second trimester. Compared with karyotype analysis, CMA/CNV-seq increased the abnormal detection rate by 10.72%, with 22q11.21 MMS being the most frequently observed pCNV type. As shown in Table 2 , only two cases of congenital heart disease were identified prenatally among six cases of 22q11.21 MMS, and one case presented no abnormality on prenatal ultrasound but was later found to have an atrial septal defect. Follow-up until age two revealed psychomotor and mental retardation. The phenotypic presentation of 22q11.21 MMS is highly variable, ranging from multiple birth defects to mild learning disabilities, with neurodevelopmental disorders and behavioral issues being the most commonly reported phenotypes [ 13 ]. Additionally, we identified a case of dupXp22.31 (1.6Mb) in which prenatal ultrasound showed fetal growth retardation. In addition to low birth weight, infants exhibit psychomotor retardation at the age of two years. Xp22.31 microduplications can lead to a variety of clinical phenotypes, including epilepsy, learning difficulties, psychomotor development retardation, and autism [ 14 ]. This study documented 24 cases of lpCNVs linked to neurocognitive disorder phenotypes, with 15q11.2 microdeletion syndrome being the most common, followed by 16p13.11 MMS. The 15q11.2 microdeletion is a newly recognized microdeletion syndrome. Furthermore, reports have shown that the 15q11.2 BP1-BP2 region is susceptible to neurological abnormalities and is associated with a series of neurological diseases, such as language and motor delays, epilepsy, and autism [ 15 ]. Additionally, the 16p13.1 region is sensitive to neurocognitive disorders, with evidence suggesting that triple-dose involvement contributes to phenotypes such as developmental delay, intellectual disabilities, learning disabilities, and behavioral abnormalities, with a reported penetrance of approximately 10.6% [ 16 ]. All 24 patients with lpCNVs continued with their pregnancies and delivered, as ultrasound examinations showed no deformities. Follow-up revealed no psychomotor retardation in these infants by the age of two. Research data in the field of pediatric genetics show that approximately 12% of children with unexplained developmental delays, intellectual disabilities, abnormal mental development, and other neurocognitive disorders may have clinically significant MMS [ 17 ]. Therefore, large-sample, multicenter studies with extended follow-ups are essential to clarify the relationship between neurodevelopmental delays in fetuses with NT thickening and CNVs associated with neurocognitive disorders. With advancements in WES technology, recent studies have reported that when ultrasound screenings indicate fetal NT thickening and rule out karyotype and microdeletion/microduplication abnormalities, continued pregnancy may result in fetal pathological deformities or developmental retardation linked to fetal monogenic diseases. Additionally, developmental retardation may be related to single-gene fetal disorders [ 18 ]. In this study, WES was conducted on 11 fetuses that excluded abnormal karyotypes and pCNV/lpCNVs, all of which presented with pathological deformities. Pathogenic or possibly pathogenic gene mutations were identified in eight cases, including two with abnormal ultrasound findings. WES confirmed single-gene disorders consistent with autosomal recessive inheritance. The remaining six cases revealed pathogenic genes associated with malformation phenotypes, indicating that genetic causes of malformation were linked to fetal NT thickening at the single-gene level before birth. Previous studies have shown that WES identified causative variants in 37.5% of fetuses with ultrasound-detected deformities and 4.8% of fetuses with isolated NT thickening [ 19 ]. Early studies have also found that approximately 9% of NT-thickened fetuses with normal chromosomes later developed deformities, with 4% experiencing long-term neurodevelopmental delay [ 20 ]. These findings suggest that even when standard karyotype and CMA analyses yield normal results, NT thickening may still be associated with neurodevelopmental disorders, even in the absence of deformities. In this study, 337 live-born infants with normal karyotypes, CNVs, and ultrasound results were followed up until two years of age. Pediatricians conducted formal clinical neurodevelopmental assessments and identified psychomotor retardation in two cases with NT measurements > 5.5 mm in the first trimester, which were confirmed as single-gene disorders via WES. Additionally, follow-ups in early childhood identified three more cases of psychomotor retardation among 44 other live-born infants. One case involved partial agenesis of the corpus callosum, and one was diagnosed with Noonan syndrome via WES. The other two cases were associated with MMS: one with del22q11.21 and the other with del22q11.31, respectively. Furthermore, WES analysis of a case with dupXp22.31 also suggested Noonan syndrome. Recent studies have emphasized that traditional karyotyping and CMA/CNV-seq testing may miss certain genetic abnormalities, particularly in fetuses without deformities. In addition, a meta-analysis suggested that in cases of fetal NT thickening with normal karyotypes and CMA, WES can identify pathogenic or potentially pathogenic genetic variants in approximately 8% of cases with fetal NT thickening, supporting the effectiveness of WES as a supplementary diagnostic method [ 21 ]. Conclusion In summary, there is a significant association between fetal NT thickening and various fetal abnormalities, although this does not provide direct evidence of fetal abnormalities. Traditional karyotyping, CMA/CNV-seq testing, and comprehensive ultrasound examinations are essential diagnostic tools for fetuses with thickened NT during the first trimester. These methods support genetic counseling and assist healthcare providers and patients in making informed decisions. More targeted WES testing should be considered to diagnose potential genetic disorders and enhance diagnostic efficiency and consultation quality, particularly in cases with structural ultrasound abnormalities or NT measurements of ≥ 5.5 mm accompanied by other abnormalities. Declarations Ethics approval and consent to participate All procedures were performed in accordance with the Declaration of Helsinki. This study protocol was approved by the Institutional Review Board of the Women and Children’s Hospital of Xiamen University. Written informed consent was obtained from all patients. Consent for publication Written informed consent was obtained from the patient for the publication of clinical details. Competing interests The authors declare no conflicts of interest. Funding This study was supported by the Xiamen Municipal Medical and Health Guidance Project (3502Z20214ZD1226). Author Contribution Mohan Wang: methodology, data curation, formal analysis, and writing – original draft; Yizhen Ji: investigation, methodology, and data curation; Yasong Xu: investigation, and formal analysis; Shiyu Sun: methodology, and formal analysis; Xiaomei Yang: software, and visualization; Li Sun: software, and visualization; Qichang Wu: conceptualization, and writing –review and editing. Acknowledgements We thank Di Shao from BGI Genomics for technical support. Data Availability The variation data and related clinical information were uploaded to the National Genomics Data Center (https://ngdc.cncb.ac.cn) with accession number GVM001199. References Bilardo CM, Timmerman E, Pajkrt E, Van Maarle M. Increased nuchal translucency in euploid fetuses—what should we be telling the parents? Prenat Diagn. 2010;30:93–102. https://doi.org/10.1002/pd.2396 . Ville Y. Nuchal translucency in the first trimester of pregnancy: ten years on and still a pain in the neck? Ultrasound Obstet Gynecol. 2001;18:5–8. https://doi.org/10.1046/j.1469-0705.2001.00483.x . Bilardo CM, Müller MA, Pajkrt E, Clur SA, Van Zalen MM, Bijlsma EK. Increased nuchal translucency thickness and normal karyotype: time for parental reassurance. Ultrasound Obstet Gyne. 2007;30:11–8. https://doi.org/10.1002/uog.4044 . Nicolaides KH, Snijders RJM, Campbell S, Gosden CM, Berry C. Ultrasonographically detectable markers of fetal chromosomal abnormalities. Lancet. 1992;340:704–7. https://doi.org/10.1016/0140-6736(92)92240-G . Atzei A, Gajewska K, Huggon IC, Allan L, Nicolaides KH. Relationship between nuchal translucency thickness and prevalence of major cardiac defects in fetuses with normal karyotype. Ultrasound Obstet Gyne. 2005;26:154–7. https://doi.org/10.1002/uog.1936 . De Domenico R, Faraci M, Hyseni E, Di Prima FAF, Valenti O, Monte S, et al. Increased nuchal traslucency in normal karyotype fetuses. J Prenat Med. 2011;5:23–6. Caughey AB, Kuppermann M, Norton ME, Washington AE. Nuchal translucency and first trimester biochemical markers for down syndrome screening: A cost-effectiveness analysis. Am J Obstet Gynecol. 2002;187:1239–45. https://doi.org/10.1067/mob.2002.127144 . Nicolaides KH, Azar G, Byrne D, Mansur C, Marks K. Fetal nuchal translucency: ultrasound screening for chromosomal defects in first trimester of pregnancy. BMJ. 1992;304:867–9. https://doi.org/10.1136/bmj.304.6831.867 . Richards S, Aziz N, Bale S, Bick D, Das S, Gastier-Foster J, et al. Standards and guidelines for the interpretation of sequence variants: a joint consensus recommendation of the American College of Medical Genetics and Genomics and the Association for Molecular Pathology. Genet Sci. 2015;17:405–24. https://doi.org/10.1038/gim.2015.30 . Study on the relationship between the fetal nuchal translucency and chromosome abnormalities. (in Chinese) Chinese Journal of Birth Health & Heredity. 2017;25:53–55 + 48. https://doi.org/10.13404/j.cnki.cjbhh.2017.03.022 Atzei A, Gajewska K, Huggon IC, Allan L, Nicolaides KH. Relationship between nuchal translucency thickness and prevalence of major cardiac defects in fetuses with normal karyotype. Ultrasound Obstet Gynecol. 2005;26:154–7. https://doi.org/10.1002/uog.1936 . Grande M, Jansen FAR, Blumenfeld YJ, Fisher A, Odibo AO, Haak MC, et al. Genomic microarray in fetuses with increased nuchal translucency and normal karyotype: a systematic review and meta-analysis. Ultrasound Obstet Gyne. 2015;46:650–8. https://doi.org/10.1002/uog.14880 . Tang KL, Antshel KM, Fremont WP, Kates WR. Behavioral and Psychiatric Phenotypes in 22q11.2 Deletion Syndrome. J Dev Behav Pediatr. 2015;36:639–50. https://doi.org/10.1097/DBP.0000000000000210 . Esplin ED, Li B, Slavotinek A, Novelli A, Battaglia A, Clark R, et al. Nine patients with Xp22.31 microduplication, cognitive deficits, seizures, and talipes anomalies. Am J Med Genet Pt A. 2014;164:2097–103. https://doi.org/10.1002/ajmg.a.36598 . Picinelli C, Lintas C, Piras IS, Gabriele S, Sacco R, Brogna C, et al. Recurrent 15q11.2 BP1-BP2 microdeletions and microduplications in the etiology of neurodevelopmental disorders. Am J Med Genet Pt B. 2016;171:1088–98. https://doi.org/10.1002/ajmg.b.32480 . Li J, Hojlo MA, Chennuri S, Gujral N, Paterson HL, Shefchek KA, et al. Underrepresentation of Phenotypic Variability of 16p13.11 Microduplication Syndrome Assessed With an Online Self-Phenotyping Tool (Phenotypr): Cohort Study. J Med Internet Res. 2021;23:e21023. https://doi.org/10.2196/21023 . Nevado J, Mergener R, Palomares-Bralo M, Souza KR, Vallespín E, Mena R, et al. New microdeletion and microduplication syndromes: a comprehensive review. Genet Mol Biol. 2014;37(1 suppl 1):210–9. https://doi.org/10.1590/S1415-47572014000200007 . Äyräs O, Eronen M, Tikkanen M, Rahkola-Soisalo P, Paavonen J, Stefanovic V. Long‐term outcome in apparently healthy children with increased nuchal translucency in the first trimester screening. Acta Obstet Gynecol Scand. 2016;95:541–6. https://doi.org/10.1111/aogs.12878 . Cao C, Liu F, Yang Y, Zhang Q, Huang J, Liu X. Prenatal whole-exome sequencing in fetuses with increased nuchal translucency. Molec Gen Gen Med. 2023;11:e2246. https://doi.org/10.1002/mgg3.2246 . Miltoft CB, Ekelund CK, Hansen BM, Lando A, Petersen OB, Skovbo P, et al. Increased nuchal translucency, normal karyotype and infant development. Ultrasound Obstet Gyne. 2012;39:28–33. https://doi.org/10.1002/uog.10060 . Di Girolamo R, Rizzo G, Khalil A, Alameddine S, Lisi G, Liberati M, et al. Whole exome sequencing in fetuses with isolated increased nuchal translucency: a systematic review and meta-analysis. J Maternal-Fetal Neonatal Med. 2023;36:2193285. https://doi.org/10.1080/14767058.2023.2193285 . Additional Declarations No competing interests reported. Cite Share Download PDF Status: Published Journal Publication published 21 Feb, 2026 Read the published version in BMC Medical Genomics → Version 1 posted Editorial decision: Revision requested 05 Jan, 2026 Reviewers agreed at journal 16 Dec, 2025 Reviews received at journal 14 Dec, 2025 Reviewers agreed at journal 07 Dec, 2025 Reviews received at journal 05 Dec, 2025 Reviewers agreed at journal 04 Dec, 2025 Reviewers agreed at journal 04 Dec, 2025 Reviewers invited by journal 04 Dec, 2025 Editor invited by journal 11 Nov, 2025 Editor assigned by journal 10 Nov, 2025 Submission checks completed at journal 10 Nov, 2025 First submitted to journal 09 Nov, 2025 You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-8072036","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":556438256,"identity":"cf4c4840-23d8-4e5f-9a0f-355aadfd1a94","order_by":0,"name":"Mohan Wang","email":"","orcid":"","institution":"Xiamen University","correspondingAuthor":false,"prefix":"","firstName":"Mohan","middleName":"","lastName":"Wang","suffix":""},{"id":556438257,"identity":"3df393cb-ddf8-4443-bae4-3166e38fed51","order_by":1,"name":"Yizhen Ji","email":"","orcid":"","institution":"Xiamen University","correspondingAuthor":false,"prefix":"","firstName":"Yizhen","middleName":"","lastName":"Ji","suffix":""},{"id":556438258,"identity":"3a80cbb6-fc5d-425c-a31c-d65986fe2d24","order_by":2,"name":"Yasong Xu","email":"","orcid":"","institution":"Xiamen University","correspondingAuthor":false,"prefix":"","firstName":"Yasong","middleName":"","lastName":"Xu","suffix":""},{"id":556438259,"identity":"19806d83-10fa-4a56-b677-34fe7d16328a","order_by":3,"name":"Shiyu Sun","email":"","orcid":"","institution":"Xiamen University","correspondingAuthor":false,"prefix":"","firstName":"Shiyu","middleName":"","lastName":"Sun","suffix":""},{"id":556438260,"identity":"3b12db4a-8115-4361-b2e7-eb589e847a3b","order_by":4,"name":"Xiaomei Yang","email":"","orcid":"","institution":"Xiamen University","correspondingAuthor":false,"prefix":"","firstName":"Xiaomei","middleName":"","lastName":"Yang","suffix":""},{"id":556438261,"identity":"65b7e70d-f324-4975-8420-f66eff36744b","order_by":5,"name":"Li Sun","email":"","orcid":"","institution":"Xiamen University","correspondingAuthor":false,"prefix":"","firstName":"Li","middleName":"","lastName":"Sun","suffix":""},{"id":556438262,"identity":"649f6e6f-12d8-42a4-96d1-4bd9c2458dba","order_by":6,"name":"Qichang Wu","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAAyUlEQVRIiWNgGAWjYDACZgY2GOvAgQ8/SNPClnhwZg9x9sC08Bgf5mDDqxIC+I4zP3vwc0ctg8GNnA+HGXgY5PnFDuDXInmYzdyw98xxoJbcDYcLLBgMZ85OwK/F4DAPmwRv2zEGM5CWGTwMCQa3idAi+ResJecBkE2kFmnethqQFgbitAD9YiYt23aAwf7MMwNgIEsQ9gvf+cPPJN+21TFItic//vDhh408vzQBLQwHwOTh+gYIV4KAcoSWOiJUjoJRMApGwYgFAKQcRaUZBnS3AAAAAElFTkSuQmCC","orcid":"","institution":"Xiamen University","correspondingAuthor":true,"prefix":"","firstName":"Qichang","middleName":"","lastName":"Wu","suffix":""}],"badges":[],"createdAt":"2025-11-10 02:38:23","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-8072036/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-8072036/v1","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.1186/s12920-026-02331-8","type":"published","date":"2026-02-21T15:57:51+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":97894432,"identity":"e2357a71-dd51-4a22-847f-c27aef171942","added_by":"auto","created_at":"2025-12-10 15:32:29","extension":"docx","order_by":0,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":261604,"visible":true,"origin":"","legend":"","description":"","filename":"ManuscriptBMCMG1106.docx","url":"https://assets-eu.researchsquare.com/files/rs-8072036/v1/555badfa334db6c7834169d9.docx"},{"id":97694829,"identity":"87812c28-ccb1-4dc9-9564-9c3ab416e049","added_by":"auto","created_at":"2025-12-08 11:26:34","extension":"json","order_by":1,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":8276,"visible":true,"origin":"","legend":"","description":"","filename":"38ebd0c752a14fd88a30e0f10f4942b1.json","url":"https://assets-eu.researchsquare.com/files/rs-8072036/v1/8784ac922d1044c0da6a5567.json"},{"id":97694832,"identity":"5f3bc725-9f88-4050-8a76-cd0753b2e91a","added_by":"auto","created_at":"2025-12-08 11:26:34","extension":"xml","order_by":2,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":97416,"visible":true,"origin":"","legend":"","description":"","filename":"38ebd0c752a14fd88a30e0f10f4942b11enriched.xml","url":"https://assets-eu.researchsquare.com/files/rs-8072036/v1/e041d301f23f7222c6479771.xml"},{"id":97694834,"identity":"1c824202-1ed1-4696-b18b-9b946f281b40","added_by":"auto","created_at":"2025-12-08 11:26:34","extension":"jpeg","order_by":3,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":206646,"visible":true,"origin":"","legend":"","description":"","filename":"floatimage1.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-8072036/v1/b2c0f70b6f309ded62e59358.jpeg"},{"id":97694835,"identity":"ee03a25a-1050-4a9b-a62f-4cf1238be9e5","added_by":"auto","created_at":"2025-12-08 11:26:35","extension":"xml","order_by":5,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":96520,"visible":true,"origin":"","legend":"","description":"","filename":"38ebd0c752a14fd88a30e0f10f4942b11structuring.xml","url":"https://assets-eu.researchsquare.com/files/rs-8072036/v1/ff8a454d1e7de5d1ad1626a0.xml"},{"id":97894110,"identity":"8a8bda32-08a4-4400-9b07-837f65b0bd7a","added_by":"auto","created_at":"2025-12-10 15:31:56","extension":"html","order_by":6,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":103057,"visible":true,"origin":"","legend":"","description":"","filename":"earlyproof.html","url":"https://assets-eu.researchsquare.com/files/rs-8072036/v1/3a178381860f11a2072490c8.html"},{"id":97694830,"identity":"dd388de7-88dd-4589-9988-b3617d8c31ec","added_by":"auto","created_at":"2025-12-08 11:26:34","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":30723,"visible":true,"origin":"","legend":"\u003cp\u003ePerinatal outcome analysis of 720 fetuses with thickened NT\u003c/p\u003e","description":"","filename":"Onlinefloatimage1.png","url":"https://assets-eu.researchsquare.com/files/rs-8072036/v1/f71898757779e36fef5a3120.png"},{"id":103252603,"identity":"8582d1df-9822-43d8-8f31-db987d6c6221","added_by":"auto","created_at":"2026-02-23 16:15:21","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1017146,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-8072036/v1/da9189b3-7e9f-440d-ba4e-6d3dc7f36bcb.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Diagnostic Value of Karyotyping, CMA/CNV-Seq, and WES in Fetuses with Thickened Nuchal Translucency: Perinatal and Two-Year Follow-Up Outcomes","fulltext":[{"header":"Key Points","content":"\u003cp\u003e1. Thickened NT linked to high chromosomal risk\u003c/p\u003e\u003cp\u003e2. CMA/CNV-seq improved detection beyond karyotype\u003c/p\u003e\u003cp\u003e3. WES showed high yield in selected cases\u003c/p\u003e"},{"header":"Background","content":"\u003cp\u003eNuchal translucency (NT) refers to fluid accumulation behind the neck of a developing fetus, which can be measured via ultrasound during early pregnancy. It serves as an important indicator of potential chromosomal abnormalities, deformities, fetal anemia, and hypoproteinemia [\u003cspan additionalcitationids=\"CR2\" citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e]. On ultrasound images, the NT appears as an echoless area behind the neck between the deep soft tissue hyperechoic band and skin hyperechoic zone [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e]. The fundamental pathophysiological mechanisms underlying NT thickening are complex and not completely understood. The primary cause of NT thickening in fetuses is associated with cardiac anatomical abnormalities. Moreover, abnormalities in the lymphatic system and reflux disorders contribute to heart failure due to congenital malformations and genetic factors [\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e]. In 1992, Nicolaides et al. first proposed the use of fetal NT thickening in the first trimester as a marker of Down syndrome [\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e]. However, the International Society of Obstetrics and Gynecology Ultrasound currently lacks a strict definition for the range of NT thickening, and the specific cut-off value for abnormal NT ranges remains controversial in various prenatal diagnostic centers, both domestically and internationally [\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e]. Most clinical practices in China currently adopt NT\u0026thinsp;\u0026ge;\u0026thinsp;2.5 mm as the cutoff.\u003c/p\u003e\u003cp\u003eRecent advances in research and the introduction of novel genetic testing technologies have revealed that NT thickening, even in the absence of chromosomal abnormalities, is associated with adverse pregnancy outcomes. These outcomes include fetal chromosomal microdeletion and microduplication syndromes (MMS), structural anomalies, certain genetic syndromes, intrauterine fetal death, and postnatal neuromotor developmental delays [\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e]. However, NT thickening is not directly correlated with fetal abnormalities. Research indicates that when fetal karyotype and microdeletion/microduplication tests yield normal results, the complete absence of deformities and growth retardation of the fetus in a systematic ultrasound examination suggests a high probability of a positive pregnancy outcome (\u0026gt;\u0026thinsp;95%) [\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e]. NT thickening and subsequent diagnostic examination are major concerns in pregnant women. The intricate association between NT and chromosomal abnormalities necessitates extensive prenatal genetic counseling and individualized diagnostic programs for pregnant women. Currently, informing pregnant women and their families about accurate fetal and pregnancy outcomes is crucial for improving their reassurance, whereas providing clinicians with reliable data to mitigate the residual risk of postpartum issues remains a major challenge in maternal-fetal medicine. However, there is no consensus on the approaches to prenatal diagnosis and management of these conditions. This study retrospectively analyzed 720 singleton cases of interventional prenatal diagnoses associated with an NT of \u0026ge;\u0026thinsp;2.5 mm in the first trimester. The analysis included copy number variations (CNV), whole-exome sequencing (WES), and fetal karyotype assessments. Additionally, this study examined fetal ultrasound screenings for deformities, tracked pregnancy outcomes, and monitored the growth and development of live infants for up to two years after birth. This study aimed to provide guidance for clinical consultation, prenatal diagnosis strategies, prognostic evaluation, and reproductive planning in cases of increased fetal NT thickness.\u003c/p\u003e"},{"header":"Materials and Methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e\u003ch2\u003eSubjects\u003c/h2\u003e\u003cp\u003eThis study retrospectively analyzed 720 singleton cases of interventional prenatal diagnosis. The cohort comprised 342 primiparous and 378 multiparous mothers who underwent prenatal examinations and ultrasonography between 11 and 13\u003csup\u003e+\u0026thinsp;6\u003c/sup\u003e weeks of pregnancy, revealing a fetal NT\u0026thinsp;\u0026ge;\u0026thinsp;2.5 mm. Data were collected from January 1, 2018, to December 31, 2021. The average age of pregnant women meeting the inclusion criteria was 31.08 (18\u0026ndash;46 years). Pregnancies associated with identified chromosomal abnormalities or significant defects diagnosed prenatally or postnatally were excluded from the analysis. In total, 213 patients underwent transabdominal chorionic villus sampling, 483 underwent transabdominal amniocentesis, and 24 underwent transabdominal umbilical cord blood sampling. All interventional prenatal diagnoses were performed, and the samples were analyzed using G-banded chromosome karyotyping and CMA/CNV-seq detection technology. Among these cases, 11 with normal fetal karyotypes and CMA/CNV-seq results were further tested using WES due to deformities. For fetuses with identified karyotypes or copy number abnormalities, couples are recommended to provide peripheral blood samples to further classify the variants as de novo or genetic. All pregnant women and their families were informed of the indications and risks associated with interventional punctures before surgery and signed an \"Informed Consent Form for Surgery.\u0026rdquo; Following the acquisition of informed consent, the study was approved by the Ethics Committee of Women and Children\u0026rsquo;s Hospital of Xiamen University.\u003c/p\u003e\u003c/div\u003e\n\u003ch3\u003eFetal NT measurement standards\u003c/h3\u003e\n\u003cp\u003eUltrasound measurement of NT thickness is based on the British Fetal Foundation Medical Society (FMF) standards for gestational ages between 11 and 13\u003csup\u003e+\u0026thinsp;6\u003c/sup\u003e weeks or when the fetal crown-rump length is approximately 45\u0026ndash;84 mm [\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e]. In this study, an NT measurement of \u0026ge;\u0026thinsp;2.5 mm was considered abnormal. High-resolution ultrasound was used, positioning the probe perpendicular to the skin of the fetal neck and back to ensure that the fetus was in a natural propensity position. A median sagittal section was obtained, and the long ribbon-like anechoic area beneath the skin of the fetal neck and back was measured using a Vernier caliper. The measurements were repeated thrice, and the maximum value was recorded as the NT thickness.\u003c/p\u003e\n\u003ch3\u003eCMA/CNV-seq detection\u003c/h3\u003e\n\u003cp\u003eIn this study, CMA was conducted in collaboration with the Beijing Beikang Medical Laboratory, and CNV-seq was performed at the BGI Genomics Medical Laboratory. All participants signed an informed consent form after a comprehensive explanation of the CMA/CNV-seq detection technology. Samples included 5\u0026ndash;10 mg of chorionic villi, 10 ml of amniotic fluid, and 2 ml of umbilical cord blood, which were sent for testing.\u003c/p\u003e\n\u003ch3\u003eWhole exome sequencing\u003c/h3\u003e\n\u003cp\u003eBriefly, 5\u0026ndash;10 mg of chorionic villi, 10 ml of amniotic fluid, and 2 ml of umbilical cord blood were collected for testing. Genomic DNA was extracted using a commercial kit. Genomic DNA was fragmented, purified, and labeled to establish the library. A MGISEQ-2000 high-throughput sequencer was used for sequencing with a PE100\u0026thinsp;+\u0026thinsp;10 sequencing type, and the original sequencing data were obtained upon completion. Sequences with HG19/HG20 reference genomes were aligned using the Burrows-Wheeler Aligner (BWA) software. The sequence capture effect was concurrently evaluated, and single nucleotide variant (SNV) and insertions/deletions (Indels) were identified using the GATK software to generate the base polymorphism results for the target region. Suspicious mutations were annotated and screened, and Sanger sequencing was performed to verify all identified pathogenic mutations, confirming the microarray capture and high-throughput sequencing results. According to the latest version of the American College of Medical Genetics and Genomics (ACMGG) genetic variation classification standards and guidelines (2015), CNVs were categorized into five groups [\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e]: benign CNVs (bCNVs), likely benign CNVs (lbCNVs), variants of uncertain significance CNVs (VOUS CNVs), likely pathogenic CNVs (lpCNVs), and pathogenic CNVs (pCNVs).\u003c/p\u003e"},{"header":"Results","content":"\u003cdiv id=\"Sec8\" class=\"Section2\"\u003e\u003ch2\u003eChromosome karyotype test results\u003c/h2\u003e\u003cp\u003eIn this study, 720 samples were subjected to G-banded chromosome karyotyping. Successful cultures were obtained from 715 cases, with 480 demonstrating normal karyotypes and 235 showing abnormal karyotypes, resulting in an abnormality rate of 32.86%. This included 211 cases with abnormal chromosome numbers: 161 cases with abnormal autosome numbers, with trisomy 21 being the most common, followed by trisomy 18 and trisomy 13. There were 50 cases of sex chromosome number abnormalities, with 45 X being the most prevalent. Additionally, 24 cases of structural chromosomal abnormalities were identified; 21 cases had unbalanced structural abnormalities, and three cases had balanced structural abnormalities (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e).\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e\u003ccaption language=\"En\"\u003e\u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e\u003cdiv class=\"CaptionContent\"\u003e\u003cp\u003eDistribution of karyotypes of 235 cases of NT thickened chromosomal abnormalities\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"2\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\"\u003e\u003cp\u003eKaryotype type\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\u003eNumber of cases\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eAbnormal chromosome number\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e211\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eAbnormal number of autosomes\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e161\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e47, XN, +\u0026thinsp;21\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e106\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e47, XN, +\u0026thinsp;18\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e42\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e47, XN, +\u0026thinsp;13\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e12\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e47, XN, +7\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e1\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eAbnormal number of sex chromosomes\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e50\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e45, X\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e40\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e47, XXX\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e2\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e47, XXY\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e5\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e47, XYY\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e3\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eChromosome structural abnormalities\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e24\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eBalance\u003c/p\u003e\u003cp\u003e46, XN, ?inv (18) (p11.3q11.2)\u003c/p\u003e\u003cp\u003e46, XN, t (1;9) (q42.1;q32)\u003c/p\u003e\u003cp\u003e46, XN, t (7;14) (q31;q13) mat,\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e3\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eImbalance\u003c/p\u003e\u003cp\u003e46, XN, del (1) (q23;q24)\u003c/p\u003e\u003cp\u003e46, XN, del (14) (q24.2q31.2)\u003c/p\u003e\u003cp\u003e46, XN, del (18) (p11.2) dn\u003c/p\u003e\u003cp\u003e46, XN, del (4) (p16.1)\u003c/p\u003e\u003cp\u003e46, XN, del (10) (q26.1) dn\u003c/p\u003e\u003cp\u003e46, XN, del (4) (q13.1q22)\u003c/p\u003e\u003cp\u003e46, XN, del (15) (q11.2q13.1)\u003c/p\u003e\u003cp\u003e46, XN, del (11) (q13.5q23.1) dn\u003c/p\u003e\u003cp\u003e46, XN, del (4) (p15.1) dn\u003c/p\u003e\u003cp\u003e46, XN, del (2) (q36.3) [\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e]/46, XN [\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e]\u003c/p\u003e\u003cp\u003e46, XN, ? dup (1) (q32q41) dn\u003c/p\u003e\u003cp\u003e46, XN, der (10) t (10;22)\u003c/p\u003e\u003cp\u003e46, XN, der (8) t (5;8) (p12;p23.1) pat\u003c/p\u003e\u003cp\u003e46, XN, der (1) t (1;4) (p36.2;q31.2) pat\u003c/p\u003e\u003cp\u003e46, XN, der (11) t (6;11) (q23;q25) mat\u003c/p\u003e\u003cp\u003e46, XN, rec (18) dup (18q) inv (18) (p11.32q12.1) mat\u003c/p\u003e\u003cp\u003e46, XN, inv (9) (p11q13), dup (11) (q24q13)\u003c/p\u003e\u003cp\u003e47, XN, +? i (12) (p10) dn\u003c/p\u003e\u003cp\u003e46, XN, i (18) (q10) [34]/46, XN, del (18) (p11.1) [\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e]\u003c/p\u003e\u003cp\u003e46, X, psu dic (Y) (q11.22 [113]/45, X [\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e]\u003c/p\u003e\u003cp\u003e47, XN, +mar [33]/46, XN [\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e] dn\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e21\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003ctfoot\u003e\u003ctr\u003e\u003ctd colspan=\"2\"\u003eNote: 47, XN, +\u0026thinsp;21 contains 4 chimerism; 47, XN, +\u0026thinsp;18 contains 1 chimerism; 47, XN, +\u0026thinsp;13 contains 1 chimerism; 45, X contains 6 chimerism; 47, XXX contains 1 chimerism.\u003c/td\u003e\u003c/tr\u003e\u003c/tfoot\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\u003c/div\u003e\n\u003ch3\u003eCMA/CNV-seq detection results\u003c/h3\u003e\n\u003cp\u003eCMA/CNV-seq successfully detected abnormalities in 211 cases with chromosomal abnormalities and 21 cases with structural chromosomal imbalances, as identified by karyotype analysis. Additionally, CMA/CNV-seq identified pCNVs in five samples in which karyotype culture failed to identify them. Among 480 cases with normal karyotypes, CMA/CNV-seq detected 23 pCNVs and 24 lpCNVs, resulting in a 10.72% increase in the detection rate compared with karyotyping. 22q11.21 MMS was the most common (26.09%) of the 23 pCNVs, followed by Yq11.223q11.23 microdeletion syndrome (17.39%) (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e). Copy number variations associated with neurocognitive disorders were detected in all 24 lpCNV cases, with 15q11.2 microdeletion syndrome being the most common (37.5%), characterized by CNVs ranging from 311.8kb to 861.33kb. Other notable CNVs included 16p13.11 MMS (12.5%) with sizes from 1.63Mb\u0026thinsp;~\u0026thinsp;3.29Mb, and 22q11.21 MMS (12.5%) ranging from 114.86kb\u0026thinsp;~\u0026thinsp;1.58Mb. Two cases of Xp22.31 micro-repeats exhibited CNV sizes of 1.6 Mb (Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e).\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab2\" border=\"1\"\u003e\u003ccaption language=\"En\"\u003e\u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e\u003cdiv class=\"CaptionContent\"\u003e\u003cp\u003ePCNVs detection in 23 fetuses with thickened NT and normal karyotype\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"5\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\"\u003e\u003cp\u003eNumber\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\u003eNT mm\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003eUltrasound results\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c4\"\u003e\u003cp\u003eCNV area and size\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c5\"\u003e\u003cp\u003eCNV involves genes and syndromes/diseases\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e4.2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eCongenital heart disease\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003edel1p36.33p36.22(10.0Mb)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003eContains 103 OMIM genes including GABRD/1p36 deletion syndrome\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e2.9\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eNo obvious structural deformity was seen\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003edel3q29q29(1.69Mb)dn\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003eContains 21 protein-coding genes including DLG1/3q29 deletion syndrome\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e3\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e3\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eNo obvious structural deformity was\u0026nbsp;seen\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003edel6p25.3p22.2(25.3Mb)\u003c/p\u003e\u003cp\u003edel6q27(2.5Mb)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003eContains 89 OMIM genes including FOXC1/6pter-p24 deletion syndrome\u003c/p\u003e\u003cp\u003eContains 14 OMIM genes including ERMARD/related to AD hereditary periventricular nodular ectopia and other diseases\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e4\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eNo obvious structural deformity was\u0026nbsp;seen\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003edel7q11.23(1.5Mb)dn\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003eContains 26 OMIM genes including ELN/related to Williams-Beuren syndrome (AD)\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e4.1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eNo obvious structural deformity was\u0026nbsp;seen\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003edel10p15.3p15.1(4.8Mb)dn\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003eContains 9 OMIM genes including ZMYND11/related to autosomal dominant intellectual disability\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e6\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e2.6\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eCongenital heart disease\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003edup10p12.2p11.23(5.7Mb)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003eContains 22 OMIM genes including ANKRD26/related to diseases such as thrombocytopenia type 2 (AD)\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e7\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e3.2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eNo obvious structural deformity was\u0026nbsp;seen\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003edel14q32.12q32.13(1.63Mb)pat\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003eContains 22 protein-coding genes/associated with pleuropulmonary blastoma\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e8\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e2.7\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eShort limbs\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eUniparental diploid (UPD) on chromosome 14\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003eContains DLK1 and other genes/related to Temple syndrome (mat) and Kagami-Ogata syndrome (pat)\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e9\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e3.1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eBilateral polycystic kidney disease\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003edel17q12(1.4Mb)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003eContains 17 OMIM genes including HNF1B/recurrent 17q12 deletion syndrome\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e10\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e2.78\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eLeft polycystic kidney, right hydronephrosis\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003edel17q12(1.5Mb)dn\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003eContains 12 OMIM genes including HNF1B/recurrent 17q12 deletion syndrome\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e11\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e2.8\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eBilateral polycystic kidney disease\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003edel17q12q12(1.99Mb)dn\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003eThis region contains 26 protein-coding genes/17q12 deletion syndrome\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e12\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e8.09\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003econgenital heart disease\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003edel22q11.21(3.1Mb)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003eContains 44 OMIM genes including TBX1/22q11.2 deletion syndrome\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e13\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e2.9\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003econgenital heart disease\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003edel22q11.21(3.1Mb)dn\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003eContains 44 OMIM genes including TBX1/22q11.2 deletion syndrome\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e14\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e4.7\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eNo obvious structural deformity was\u0026nbsp;seen\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003edel22q11.21(2.8Mb)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003eContains 42 OMIM genes including TBX1/22q11.2 deletion syndrome\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e15\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e2.9\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eNo obvious structural deformity was\u0026nbsp;seen\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003edel22q11.21q11.23(2.6Mb)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003eContains 30 OMIM genes including BCR/22q11.2 distal deletion syndrome\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e16\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e3.37\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eNo obvious structural deformity was\u0026nbsp;seen\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003edup22q11.21(2.8Mb)dn\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003eContains 41 OMIM genes including CLTCL1/22q11.2 microduplication syndrome\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e17\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e3.94\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eNo obvious structural deformity was\u0026nbsp;seen\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003edup22q11.21q11.21(2.25Mb)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003eThis region contains 37 protein-coding genes/22q11.2 duplication syndrome\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e18\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e2.8\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eIntrauterine growth retardation\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003edupXp22.31(1.6Mb)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003eContains 4 OMIM genes including STS/related to clinical phenotypes such as ichthyosis, global developmental delay, autism, and epilepsy\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e19\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e7.82\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eNeck hygroma, generalized edema\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003edelXp22.33q28(155.27Mb)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003eAssociated with Turner syndrome\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e20\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e5.7\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eNeck hygroma, generalized edema\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003edelYq11.223q11.23(2.25Mb)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003eContains 17 protein-coding genes including AZFc and DAZ regions/Y-linked spermatogenesis disorder\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e21\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e3.49\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eNo obvious structural deformity was seen\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003edelYq11.223q11.23(3.11Mb)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003eThis region contains 13 protein-coding genes/Y-linked spermatogenesis disorders.\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e22\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e3\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eNo obvious structural deformity was seen\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003edelYq11.223q11.23(2.16Mb)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003eContains 7 protein-coding genes including BPY2/Y-linked spermatogenesis disorder\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e23\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e3.1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eNo obvious structural deformity was seen\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003edelYq11.223q11.23(3.55Mb) pat\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003eContains 10 protein-coding genes including BPY2/Y-linked spermatogenesis disorder\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab3\" border=\"1\"\u003e\u003ccaption language=\"En\"\u003e\u003cdiv class=\"CaptionNumber\"\u003eTable 3\u003c/div\u003e\u003cdiv class=\"CaptionContent\"\u003e\u003cp\u003eDetection results of LPCNVs in 7 fetuses with thickened NT and normal karyotype\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"4\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\"\u003e\u003cp\u003eNumber\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\u003eNTmm\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003eCNV area and size\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c4\"\u003e\u003cp\u003eCNV involves genes and syndromes/diseases\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e6.4\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003edup4q22.1(1.8Mb)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eContains 3 OMIM genes including SNCA/related to diseases such as dementia with Lewy bodies (AD)\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e3.2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003edel5p15.33p15.33(1.05Mb)dn\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e18 protein-coding genes, including the SDHA gene, are related to diseases such as delayed optic atrophy (AD)\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e3\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e4\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003edup5p15.1p14.1(8.6Mb)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eContains 5 OMIM genes including CDH18/related to clinical phenotypes such as autism, global developmental delay, and epilepsy\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e4\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e4.3\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003edup6q25.3(2.4Mb)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eContains 10 OMIM genes including ARID1B/related to CoffinSiris syndrome (AD) disease, and clinical phenotypes include intellectual disability, alopecia, sensorineural deafness, etc.\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e2.9\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003edup7p21.3(3.9Mb)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eContains TMEM106B and other 5 OMIM genes/related to hypomyelination leukodystrophy 16 (AD) disease, clinical phenotypes include intellectual disability, learning disabilities, epileptic seizures, gait ataxia, etc.\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e6\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e3.24\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003edel20p12.3(1.4Mb)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eInvolving PLCB1 and other 3 OMIM genes/associated with early infantile epileptic encephalopathy type 12 (AR)\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e7\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e3.3\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003edelXq28(376.0Kb)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eContaining part of the AFF2 gene segment/FRAXE type intellectual disability (X-linked recessive), clinical phenotypes include mild to moderate intellectual disability, cognitive difficulties, communication disorders, ADHD, autistic behavior, etc.\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\n\u003ch3\u003eExcluding the correlation between pathogenic chromosomal karyotypic abnormalities and pCNVs/lpCNVs, NT thickening and pathological deformities\u003c/h3\u003e\n\u003cp\u003eIn this study, pathogenic chromosomal karyotype abnormalities and pCNV/lpCNVs were excluded, and 85 cases of NT-thickened fetal ultrasound examinations were found to have pathological deformities (19.6%). Among them, 24 presented with fetal neck cystic hygroma, generalized skin edema, and bilateral pleural effusion. The remaining 61 patients had deformities, including 17 with multiple malformations and 44 with single malformations. Systemic deformities were classified as follows: 25 cases of cardiac malformations, 19 cases of skeletal system deformities, and 18 cases of thoracic and abdominal deformities, including eight cases of omphalocele, seven cases of gastroschisis, one case of diaphragmatic hernia, one case of duodenal atresia, and one case of an abdominal mass. Additionally, eight cases of renal dysplasia and seven cases of abnormal nervous system development were identified. Cardiac malformations were the most frequently observed structural abnormality. Among the 11 fetuses with pathological deformities tested using WES, eight were found to have pathogenic or possibly pathogenic gene mutations, resulting in a detection rate of 72.7%. Two patients had similar abnormal ultrasound results on prior examinations. WES identified monogenic diseases consistent with autosomal recessive inheritance. Ultrasonography in three cases showed abnormal skeletal development, and pathogenic genes associated with skeletal dysplasia were detected. The remaining three cases exhibited pathogenic genes linked to abnormal phenotypes (Table\u0026nbsp;\u003cspan refid=\"Tab4\" class=\"InternalRef\"\u003e4\u003c/span\u003e).\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab4\" border=\"1\"\u003e\u003ccaption language=\"En\"\u003e\u003cdiv class=\"CaptionNumber\"\u003eTable 4\u003c/div\u003e\u003cdiv class=\"CaptionContent\"\u003e\u003cp\u003eThe karyotype and CMA/CNV-seq of 8 fetal cases with thickened NT were normal and WES detected pathogenic/suspected pathogenic variants\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"5\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\"\u003e\u003cp\u003eNumber\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\u003eNT mm\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003eUltrasound test results\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c4\"\u003e\u003cp\u003eAdverse reproductive history\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c5\"\u003e\u003cp\u003eMutated genes/associated diseases/inheritance patterns\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e3.4\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eShort limb deformity, narrow chest and large abdomen\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eNone\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003eFGFR3:c.2420G\u0026thinsp;\u0026gt;\u0026thinsp;T(p.807Leufs*?) dn Lethal osteoplasia/AD\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e12.7\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eGeneralized skin edema and bone dysplasia\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eNone\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003eCOL2A1:c.3022G\u0026thinsp;\u0026gt;\u0026thinsp;C (p.Gly1008Arg)dn osteoarthritis with mild achondroplasia/AD\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e3\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e5.4\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eNeck hygroma, systemic skin edema, short limbs, abnormal intracardiac structure\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eNone\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003eSLC26A2:c.2018A\u0026thinsp;\u0026gt;\u0026thinsp;T(p.Asp673Val) heterozygous (inherited from father)\u003c/p\u003e\u003cp\u003eSLC26A2:c.2046G\u0026thinsp;\u0026gt;\u0026thinsp;A(p.Leu682Leu) heterozygous (inherited from mother)\u003c/p\u003e\u003cp\u003eDeformative bone dysplasia, chondrogenesis type 1B/AR\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e4\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e4.79\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eOmphalocele, ventricular septal defect\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eSimilar ultrasound abnormalities recur\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003ePIGN:c.963G\u0026thinsp;\u0026gt;\u0026thinsp;A(p.Gln321Gln)\u003c/p\u003e\u003cp\u003eThe fetus is homozygous (inherited from both father and mother)\u003c/p\u003e\u003cp\u003eMultiple congenital malformations-hypotonia-epilepsy syndrome type 1/AR\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e7.35\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eGeneralized skin edema\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003ePast unexplained neonatal death\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003eLZTR1:c.742G\u0026thinsp;\u0026gt;\u0026thinsp;A(p.Gly248Arg) (inherited from mother)\u003c/p\u003e\u003cp\u003eNoonan syndrome/AD\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e6\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e6.9\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eGeneralized skin edema\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eSimilar ultrasound abnormalities occur again\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003eCHRNA1:c.1128delG(p.Pro377Leufs*10) heterozygous (inherited from mother)\u003c/p\u003e\u003cp\u003eCHRNA1:c.505T\u0026thinsp;\u0026gt;\u0026thinsp;C(p.Trp169Arg) heterozygous (inherited from father)\u003c/p\u003e\u003cp\u003eMultiple fatal pterygium syndrome/AR\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e7\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e9\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eGeneralized skin edema\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eNone\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003eKMT2D:c.9769A\u0026thinsp;\u0026gt;\u0026thinsp;T(p.Lys3257*)dnKabuki syndrome type 1/AD\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e8\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e6.98\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eOmphalocele, general skin edema\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eNone\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003eMAP2K2:c.1073_1074dupCG(p.Asp359Argfs*3)dncardiofacial-cutaneous syndrome type 4/AD\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\u003cdiv id=\"Sec11\" class=\"Section2\"\u003e\u003ch2\u003eAnalysis of perinatal outcomes of 720 cases of fetal NT thickening\u003c/h2\u003e\u003cp\u003eAmong the 720 singleton pregnancies, there were 339 pregnancy terminations and 381 live births (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e).\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003eIn this study, 211 cases of chromosomal abnormalities and 20 cases of structural chromosomal imbalances diagnosed via karyotype analysis were terminated, accounting for 68.1% of terminations. Additionally, 71 cases without karyotype abnormalities or pCNV/lpCNVs but with pathological deformities detected via ultrasound accounted for 20.9% of the cases. Furthermore, 21 cases of pCNVs (including five cases in which karyotype culture failed) and five cases of lpCNVs detected by CMA/CNV-seq led to pregnancy termination (7.7%). Eleven pregnancies with normal fetal karyotypes, CNVs, and ultrasound results were terminated, accounting for 3.2% of all pregnancies. Among these, five cases were miscarriages, one was a stillbirth, three were diagnosed with severe α-thalassemia, and two cases of premature intrauterine growth restriction and abnormal umbilical blood flow required termination.\u003c/p\u003e\u003cp\u003eAmong the 381 live births, 337 had normal fetal karyotypes, CNVs, and ultrasound findings. Of these, 317 were full-term deliveries and 20 were premature births. The birth weight of these newborns was 3367.0\u0026thinsp;\u0026plusmn;\u0026thinsp;450.6 g, resulting in a live birth rate of 96.8%. The remaining 44 live births included 14 cases of pathological deformities detected via ultrasound, excluding karyotype abnormalities and pCNVs/lpCNVs. There were also 19 cases of lpCNVs, three cases of balanced chromosomal translocations with no ultrasound abnormalities, seven cases of pCNVs, and one case of unbalanced chromosomal translocation.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec12\" class=\"Section2\"\u003e\u003ch2\u003eAnalysis of the follow-up status of 381 live-born infants in early childhood\u003c/h2\u003e\u003cp\u003eAll live births were followed up in early childhood within the first two years, during which a pediatrician performed clinical neurodevelopmental assessments. Five cases of psychomotor retardation were identified. One case involved del22q11.21 (2.8Mb), which exhibited no obvious deformities on prenatal ultrasound but was later diagnosed with congenital heart disease (atrial septal defect) after birth. By two years of age, the child exhibited psychomotor retardation and mental impairments. Another case involved dupXp22.31 (1.6Mb), with prenatal ultrasound showing fetal growth retardation and polyhydramnios. This infant was born full-term via vaginal delivery with a birth weight of 2050 g, and additional WES testing confirmed Noonan syndrome. In another case, prenatal ultrasound suggested a possible partial absence of the corpus callosum, which was later confirmed by ultrasound at six months post-birth, alongside a diagnosis of congenital heart disease (atrial septal defect). WES also suggested Noonan syndrome in this case. Among the 337 live births with normal karyotypes, CNVs, and ultrasound results, one case had an NT measurement of 8.69 mm in the first trimester. WES testing identified a heterozygous mutation (dn) in SETD5 associated with autosomal dominant intellectual developmental disorder-23 (MRD23). Another case, with an NT measurement of 9.1 mm in the first trimester, was found to have a SOS1 gene mutation (dn) associated with Noonan syndrome type 4.\u003c/p\u003e\u003c/div\u003e"},{"header":"Discussion","content":"\u003cp\u003eThis study involved 720 pregnant women with first-trimester NT measurements\u0026thinsp;\u0026ge;\u0026thinsp;2.5 mm, each undergoing interventional prenatal diagnosis with follow-up during the perinatal period and early childhood (up to 2 years after birth). The results confirm that early fetal NT thickening is associated with adverse pregnancy outcomes, such as chromosomal abnormalities, chromosomal MMS, structural fetal anomalies, specific genetic syndromes, intrauterine fetal death, and postnatal neuromotor development delays. In this study, the rate of fetal karyotype abnormalities was as high as 32.86%, with trisomy 21 being the most common, highlighting a strong association between fetal NT thickening and chromosomal abnormalities, particularly in Down syndrome. A domestic study using a 2.5 mm NT threshold and amniotic fluid karyotype analysis of 4028 pregnant women reported a chromosomal abnormality rate of 22.02% among fetuses with NT thickening, which was significantly higher than that in fetuses without NT thickening, with trisomy 21 being the most prevalent [\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eMoreover, even in the absence of chromosomal abnormalities, fetuses with NT thickening face heightened risks of a wide range of congenital anomalies, including isolated structural defects, genetic syndromes, and neurodevelopmental delays that often manifest in early childhood [\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e]. A Meta-analysis revealed that CMA technology could detect an additional 5% of pathogenic variants in cases of fetal NT thickening, with 22q11.2 MMS, 10q26.12q26.3 deletions, and 12q21q22 deletions identified as the most frequent pathogenic CNVs [\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e]. Our results confirmed that after excluding chromosomal karyotype abnormalities, fetal NT thickening was associated with MMS in the second trimester. Compared with karyotype analysis, CMA/CNV-seq increased the abnormal detection rate by 10.72%, with 22q11.21 MMS being the most frequently observed pCNV type. As shown in Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e, only two cases of congenital heart disease were identified prenatally among six cases of 22q11.21 MMS, and one case presented no abnormality on prenatal ultrasound but was later found to have an atrial septal defect. Follow-up until age two revealed psychomotor and mental retardation. The phenotypic presentation of 22q11.21 MMS is highly variable, ranging from multiple birth defects to mild learning disabilities, with neurodevelopmental disorders and behavioral issues being the most commonly reported phenotypes [\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eAdditionally, we identified a case of dupXp22.31 (1.6Mb) in which prenatal ultrasound showed fetal growth retardation. In addition to low birth weight, infants exhibit psychomotor retardation at the age of two years. Xp22.31 microduplications can lead to a variety of clinical phenotypes, including epilepsy, learning difficulties, psychomotor development retardation, and autism [\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e]. This study documented 24 cases of lpCNVs linked to neurocognitive disorder phenotypes, with 15q11.2 microdeletion syndrome being the most common, followed by 16p13.11 MMS. The 15q11.2 microdeletion is a newly recognized microdeletion syndrome. Furthermore, reports have shown that the 15q11.2 BP1-BP2 region is susceptible to neurological abnormalities and is associated with a series of neurological diseases, such as language and motor delays, epilepsy, and autism [\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eAdditionally, the 16p13.1 region is sensitive to neurocognitive disorders, with evidence suggesting that triple-dose involvement contributes to phenotypes such as developmental delay, intellectual disabilities, learning disabilities, and behavioral abnormalities, with a reported penetrance of approximately 10.6% [\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e]. All 24 patients with lpCNVs continued with their pregnancies and delivered, as ultrasound examinations showed no deformities. Follow-up revealed no psychomotor retardation in these infants by the age of two. Research data in the field of pediatric genetics show that approximately 12% of children with unexplained developmental delays, intellectual disabilities, abnormal mental development, and other neurocognitive disorders may have clinically significant MMS [\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e]. Therefore, large-sample, multicenter studies with extended follow-ups are essential to clarify the relationship between neurodevelopmental delays in fetuses with NT thickening and CNVs associated with neurocognitive disorders.\u003c/p\u003e\u003cp\u003eWith advancements in WES technology, recent studies have reported that when ultrasound screenings indicate fetal NT thickening and rule out karyotype and microdeletion/microduplication abnormalities, continued pregnancy may result in fetal pathological deformities or developmental retardation linked to fetal monogenic diseases. Additionally, developmental retardation may be related to single-gene fetal disorders [\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e]. In this study, WES was conducted on 11 fetuses that excluded abnormal karyotypes and pCNV/lpCNVs, all of which presented with pathological deformities. Pathogenic or possibly pathogenic gene mutations were identified in eight cases, including two with abnormal ultrasound findings. WES confirmed single-gene disorders consistent with autosomal recessive inheritance. The remaining six cases revealed pathogenic genes associated with malformation phenotypes, indicating that genetic causes of malformation were linked to fetal NT thickening at the single-gene level before birth. Previous studies have shown that WES identified causative variants in 37.5% of fetuses with ultrasound-detected deformities and 4.8% of fetuses with isolated NT thickening [\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eEarly studies have also found that approximately 9% of NT-thickened fetuses with normal chromosomes later developed deformities, with 4% experiencing long-term neurodevelopmental delay [\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e]. These findings suggest that even when standard karyotype and CMA analyses yield normal results, NT thickening may still be associated with neurodevelopmental disorders, even in the absence of deformities. In this study, 337 live-born infants with normal karyotypes, CNVs, and ultrasound results were followed up until two years of age. Pediatricians conducted formal clinical neurodevelopmental assessments and identified psychomotor retardation in two cases with NT measurements\u0026thinsp;\u0026gt;\u0026thinsp;5.5 mm in the first trimester, which were confirmed as single-gene disorders via WES. Additionally, follow-ups in early childhood identified three more cases of psychomotor retardation among 44 other live-born infants. One case involved partial agenesis of the corpus callosum, and one was diagnosed with Noonan syndrome via WES. The other two cases were associated with MMS: one with del22q11.21 and the other with del22q11.31, respectively. Furthermore, WES analysis of a case with dupXp22.31 also suggested Noonan syndrome. Recent studies have emphasized that traditional karyotyping and CMA/CNV-seq testing may miss certain genetic abnormalities, particularly in fetuses without deformities. In addition, a meta-analysis suggested that in cases of fetal NT thickening with normal karyotypes and CMA, WES can identify pathogenic or potentially pathogenic genetic variants in approximately 8% of cases with fetal NT thickening, supporting the effectiveness of WES as a supplementary diagnostic method [\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e].\u003c/p\u003e"},{"header":"Conclusion","content":"\u003cp\u003eIn summary, there is a significant association between fetal NT thickening and various fetal abnormalities, although this does not provide direct evidence of fetal abnormalities. Traditional karyotyping, CMA/CNV-seq testing, and comprehensive ultrasound examinations are essential diagnostic tools for fetuses with thickened NT during the first trimester. These methods support genetic counseling and assist healthcare providers and patients in making informed decisions. More targeted WES testing should be considered to diagnose potential genetic disorders and enhance diagnostic efficiency and consultation quality, particularly in cases with structural ultrasound abnormalities or NT measurements of \u0026ge;\u0026thinsp;5.5 mm accompanied by other abnormalities.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003ch2\u003eEthics approval and consent to participate\u003c/h2\u003e\u003cp\u003e All procedures were performed in accordance with the Declaration of Helsinki. This study protocol was approved by the Institutional Review Board of the Women and Children\u0026rsquo;s Hospital of Xiamen University. Written informed consent was obtained from all patients.\u003c/p\u003e\u003c/p\u003e\u003cp\u003e\u003cstrong\u003eConsent for publication\u003c/strong\u003e\u003cp\u003e Written informed consent was obtained from the patient for the publication of clinical details.\u003c/p\u003e\u003c/p\u003e\u003cp\u003e\u003ch2\u003eCompeting interests\u003c/h2\u003e\u003cp\u003eThe authors declare no conflicts of interest.\u003c/p\u003e\u003c/p\u003e\u003ch2\u003eFunding\u003c/h2\u003e\u003cp\u003eThis study was supported by the Xiamen Municipal Medical and Health Guidance Project (3502Z20214ZD1226).\u003c/p\u003e\u003ch2\u003eAuthor Contribution\u003c/h2\u003e\u003cp\u003eMohan Wang: methodology, data curation, formal analysis, and writing \u0026ndash; original draft; Yizhen Ji: investigation, methodology, and data curation; Yasong Xu: investigation, and formal analysis; Shiyu Sun: methodology, and formal analysis; Xiaomei Yang: software, and visualization; Li Sun: software, and visualization; Qichang Wu: conceptualization, and writing \u0026ndash;review and editing.\u003c/p\u003e\u003ch2\u003eAcknowledgements\u003c/h2\u003e\u003cp\u003eWe thank Di Shao from BGI Genomics for technical support.\u003c/p\u003e\u003ch2\u003eData Availability\u003c/h2\u003e\u003cp\u003eThe variation data and related clinical information were uploaded to the National Genomics Data Center (https://ngdc.cncb.ac.cn) with accession number GVM001199.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eBilardo CM, Timmerman E, Pajkrt E, Van Maarle M. Increased nuchal translucency in euploid fetuses\u0026mdash;what should we be telling the parents? Prenat Diagn. 2010;30:93\u0026ndash;102. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1002/pd.2396\u003c/span\u003e\u003cspan address=\"10.1002/pd.2396\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eVille Y. Nuchal translucency in the first trimester of pregnancy: ten years on and still a pain in the neck? Ultrasound Obstet Gynecol. 2001;18:5\u0026ndash;8. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1046/j.1469-0705.2001.00483.x\u003c/span\u003e\u003cspan address=\"10.1046/j.1469-0705.2001.00483.x\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eBilardo CM, M\u0026uuml;ller MA, Pajkrt E, Clur SA, Van Zalen MM, Bijlsma EK. Increased nuchal translucency thickness and normal karyotype: time for parental reassurance. Ultrasound Obstet Gyne. 2007;30:11\u0026ndash;8. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1002/uog.4044\u003c/span\u003e\u003cspan address=\"10.1002/uog.4044\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eNicolaides KH, Snijders RJM, Campbell S, Gosden CM, Berry C. Ultrasonographically detectable markers of fetal chromosomal abnormalities. Lancet. 1992;340:704\u0026ndash;7. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/0140-6736(92)92240-G\u003c/span\u003e\u003cspan address=\"10.1016/0140-6736(92)92240-G\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eAtzei A, Gajewska K, Huggon IC, Allan L, Nicolaides KH. Relationship between nuchal translucency thickness and prevalence of major cardiac defects in fetuses with normal karyotype. Ultrasound Obstet Gyne. 2005;26:154\u0026ndash;7. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1002/uog.1936\u003c/span\u003e\u003cspan address=\"10.1002/uog.1936\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eDe Domenico R, Faraci M, Hyseni E, Di Prima FAF, Valenti O, Monte S, et al. Increased nuchal traslucency in normal karyotype fetuses. J Prenat Med. 2011;5:23\u0026ndash;6.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eCaughey AB, Kuppermann M, Norton ME, Washington AE. Nuchal translucency and first trimester biochemical markers for down syndrome screening: A cost-effectiveness analysis. Am J Obstet Gynecol. 2002;187:1239\u0026ndash;45. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1067/mob.2002.127144\u003c/span\u003e\u003cspan address=\"10.1067/mob.2002.127144\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eNicolaides KH, Azar G, Byrne D, Mansur C, Marks K. Fetal nuchal translucency: ultrasound screening for chromosomal defects in first trimester of pregnancy. BMJ. 1992;304:867\u0026ndash;9. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1136/bmj.304.6831.867\u003c/span\u003e\u003cspan address=\"10.1136/bmj.304.6831.867\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eRichards S, Aziz N, Bale S, Bick D, Das S, Gastier-Foster J, et al. Standards and guidelines for the interpretation of sequence variants: a joint consensus recommendation of the American College of Medical Genetics and Genomics and the Association for Molecular Pathology. Genet Sci. 2015;17:405\u0026ndash;24. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1038/gim.2015.30\u003c/span\u003e\u003cspan address=\"10.1038/gim.2015.30\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eStudy on the relationship between the fetal nuchal translucency and chromosome abnormalities. (in Chinese) Chinese Journal of Birth Health \u0026amp; Heredity. 2017;25:53\u0026ndash;55\u0026thinsp;+\u0026thinsp;48. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.13404/j.cnki.cjbhh.2017.03.022\u003c/span\u003e\u003cspan address=\"10.13404/j.cnki.cjbhh.2017.03.022\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eAtzei A, Gajewska K, Huggon IC, Allan L, Nicolaides KH. Relationship between nuchal translucency thickness and prevalence of major cardiac defects in fetuses with normal karyotype. Ultrasound Obstet Gynecol. 2005;26:154\u0026ndash;7. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1002/uog.1936\u003c/span\u003e\u003cspan address=\"10.1002/uog.1936\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eGrande M, Jansen FAR, Blumenfeld YJ, Fisher A, Odibo AO, Haak MC, et al. Genomic microarray in fetuses with increased nuchal translucency and normal karyotype: a systematic review and meta-analysis. Ultrasound Obstet Gyne. 2015;46:650\u0026ndash;8. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1002/uog.14880\u003c/span\u003e\u003cspan address=\"10.1002/uog.14880\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eTang KL, Antshel KM, Fremont WP, Kates WR. Behavioral and Psychiatric Phenotypes in 22q11.2 Deletion Syndrome. J Dev Behav Pediatr. 2015;36:639\u0026ndash;50. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1097/DBP.0000000000000210\u003c/span\u003e\u003cspan address=\"10.1097/DBP.0000000000000210\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eEsplin ED, Li B, Slavotinek A, Novelli A, Battaglia A, Clark R, et al. Nine patients with Xp22.31 microduplication, cognitive deficits, seizures, and talipes anomalies. Am J Med Genet Pt A. 2014;164:2097\u0026ndash;103. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1002/ajmg.a.36598\u003c/span\u003e\u003cspan address=\"10.1002/ajmg.a.36598\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003ePicinelli C, Lintas C, Piras IS, Gabriele S, Sacco R, Brogna C, et al. Recurrent 15q11.2 BP1-BP2 microdeletions and microduplications in the etiology of neurodevelopmental disorders. Am J Med Genet Pt B. 2016;171:1088\u0026ndash;98. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1002/ajmg.b.32480\u003c/span\u003e\u003cspan address=\"10.1002/ajmg.b.32480\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eLi J, Hojlo MA, Chennuri S, Gujral N, Paterson HL, Shefchek KA, et al. Underrepresentation of Phenotypic Variability of 16p13.11 Microduplication Syndrome Assessed With an Online Self-Phenotyping Tool (Phenotypr): Cohort Study. J Med Internet Res. 2021;23:e21023. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.2196/21023\u003c/span\u003e\u003cspan address=\"10.2196/21023\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eNevado J, Mergener R, Palomares-Bralo M, Souza KR, Vallesp\u0026iacute;n E, Mena R, et al. New microdeletion and microduplication syndromes: a comprehensive review. Genet Mol Biol. 2014;37(1 suppl 1):210\u0026ndash;9. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1590/S1415-47572014000200007\u003c/span\u003e\u003cspan address=\"10.1590/S1415-47572014000200007\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003e\u0026Auml;yr\u0026auml;s O, Eronen M, Tikkanen M, Rahkola-Soisalo P, Paavonen J, Stefanovic V. Long‐term outcome in apparently healthy children with increased nuchal translucency in the first trimester screening. Acta Obstet Gynecol Scand. 2016;95:541\u0026ndash;6. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1111/aogs.12878\u003c/span\u003e\u003cspan address=\"10.1111/aogs.12878\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eCao C, Liu F, Yang Y, Zhang Q, Huang J, Liu X. Prenatal whole-exome sequencing in fetuses with increased nuchal translucency. Molec Gen Gen Med. 2023;11:e2246. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1002/mgg3.2246\u003c/span\u003e\u003cspan address=\"10.1002/mgg3.2246\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eMiltoft CB, Ekelund CK, Hansen BM, Lando A, Petersen OB, Skovbo P, et al. Increased nuchal translucency, normal karyotype and infant development. Ultrasound Obstet Gyne. 2012;39:28\u0026ndash;33. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1002/uog.10060\u003c/span\u003e\u003cspan address=\"10.1002/uog.10060\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eDi Girolamo R, Rizzo G, Khalil A, Alameddine S, Lisi G, Liberati M, et al. Whole exome sequencing in fetuses with isolated increased nuchal translucency: a systematic review and meta-analysis. J Maternal-Fetal Neonatal Med. 2023;36:2193285. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1080/14767058.2023.2193285\u003c/span\u003e\u003cspan address=\"10.1080/14767058.2023.2193285\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"bmc-medical-genomics","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"mgnm","sideBox":"Learn more about [BMC Medical Genomics](http://bmcmedgenomics.biomedcentral.com/)","snPcode":"","submissionUrl":"https://www.editorialmanager.com/mgnm/default.aspx","title":"BMC Medical Genomics","twitterHandle":"BMC_series","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"em","reportingPortfolio":"BMC Series","inReviewEnabled":true,"inReviewRevisionsEnabled":true},"keywords":"Nuchal translucency, Chromosome karyotype, CMA/CNV-seq, Perinatal outcomes","lastPublishedDoi":"10.21203/rs.3.rs-8072036/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-8072036/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003e\u003cstrong\u003eBackground\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis study aimed to analyze the perinatal and pediatric outcomes of fetuses with thickened nuchal translucency (NT ≥ 2.5 mm) to enhance prenatal diagnostic strategies.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eStudy Design\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eA total of 720 pregnant women with NT ≥ 2.5 mm in the first trimester underwent interventional prenatal diagnosis. These participants were followed up during the perinatal and pediatric periods (2 years after birth).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eResults\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe incidence of fetal chromosomal karyotype abnormalities was 32.86%, with trisomy 21 being the most common abnormality. Chromosomal microarray analysis (CMA) and copy number variation sequencing (CNV-seq) increased the detection rate by 10.72%. The most prevalent pathogenic copy number variations (pCNVs) and likely pathogenic copy number variations (lpCNVs) were associated with 22q11.21 microdeletion/duplication syndrome and 15q11.2 microdeletion syndrome, respectively. Excluding pathogenic karyotype abnormalities and pCNV/lpCNVs, the rate of pathological deformities was 19.6%. Whole-exome sequencing (WES) was performed in 11 cases, yielding a detection rate of 72.7%. There were 339 pregnancy terminations and 381 live births, of which 337 had normal karyotypes, CNVs, and ultrasound results, resulting in a live-birth incidence of 96.8%. In addition, two cases of psychomotor retardation were identified.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConclusion\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eTraditional karyotyping, CMA/CNV-seq testing, and detailed ultrasound examinations are vital diagnostic tools that support genetic counseling for fetuses with thickened NT in the first trimester. Therefore, novel and efficient WES testing is required.\u003c/p\u003e","manuscriptTitle":"Diagnostic Value of Karyotyping, CMA/CNV-Seq, and WES in Fetuses with Thickened Nuchal Translucency: Perinatal and Two-Year Follow-Up Outcomes","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-12-08 11:26:30","doi":"10.21203/rs.3.rs-8072036/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2026-01-05T09:39:37+00:00","index":"","fulltext":""},{"type":"reviewerAgreed","content":"181660401762929717536301870152417865932","date":"2025-12-16T12:05:22+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-12-14T12:10:57+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"226409873672123389588870261591662467103","date":"2025-12-07T09:53:17+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-12-05T08:51:33+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"45537351613849537366714607510516169397","date":"2025-12-04T13:40:30+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"330985108618856929964335127319524767785","date":"2025-12-04T12:30:11+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2025-12-04T11:44:34+00:00","index":"","fulltext":""},{"type":"editorInvited","content":"","date":"2025-11-11T05:34:55+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2025-11-10T13:03:28+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2025-11-10T13:01:40+00:00","index":"","fulltext":""},{"type":"submitted","content":"BMC Medical Genomics","date":"2025-11-10T02:36:04+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"bmc-medical-genomics","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"mgnm","sideBox":"Learn more about [BMC Medical Genomics](http://bmcmedgenomics.biomedcentral.com/)","snPcode":"","submissionUrl":"https://www.editorialmanager.com/mgnm/default.aspx","title":"BMC Medical Genomics","twitterHandle":"BMC_series","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"em","reportingPortfolio":"BMC Series","inReviewEnabled":true,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"68f7e91e-8264-4738-8ec0-9d815a8522ce","owner":[],"postedDate":"December 8th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[],"tags":[],"updatedAt":"2026-02-23T16:14:19+00:00","versionOfRecord":{"articleIdentity":"rs-8072036","link":"https://doi.org/10.1186/s12920-026-02331-8","journal":{"identity":"bmc-medical-genomics","isVorOnly":false,"title":"BMC Medical Genomics"},"publishedOn":"2026-02-21 15:57:51","publishedOnDateReadable":"February 21st, 2026"},"versionCreatedAt":"2025-12-08 11:26:30","video":"","vorDoi":"10.1186/s12920-026-02331-8","vorDoiUrl":"https://doi.org/10.1186/s12920-026-02331-8","workflowStages":[]},"version":"v1","identity":"rs-8072036","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-8072036","identity":"rs-8072036","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}