Prenatal genetic evaluation and outcomes in pregnancies with first-occurrence typical orofacial clefts

Prenatal genetic evaluation and outcomes in pregnancies with first-occurrence typical orofacial clefts | 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 Prenatal genetic evaluation and outcomes in pregnancies with first-occurrence typical orofacial clefts Xiaoqing Wu, Xiaorui Xie, Jinzhou Lu, Linjuan Su, Bin Liang, Na Lin, and 3 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-6191104/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Background Orofacial clefts (OFCs) are among the most common birth defects. This study aimed to investigate the prenatal genetic evaluation and pregnancy outcomes of pregnancies with first-occurrence typical OFCs. Methods We retrospectively reviewed 205 first-occurrence OFCs pregnancies during 2010 and 2024, including cleft lip (CL, n = 42), cleft palate (CP, n = 31), and cleft lip and palate (CLP, n = 132). Based on the presence of additional malformations, cases were categorized as isolated (n = 153) or non-isolated (n = 52). Conventional karyotyping was used to detect chromosomal abnormalities, and single nucleotide polymorphism array (SNP array) analysis was performed in 138 cases to identify submicroscopic aberrations. Results The proportions of isolated cases for CL, CP, and CLP were 95.2% (40/42), 83.8% (26/31), and 65.9% (87/132), respectively. Conventional karyotyping identified chromosomal abnormalities in 24 cases (11.7%), with trisomy 13 being the most common (12 cases, 50.0%), followed by trisomy 18 (5 cases, 20.8%). All abnormalities were observed in the non-isolated group, where the chromosomal abnormality rate was 46.2% (24/52). In this group, the chromosomal abnormality rates for CL, CP, and CLP were 50.0% (1/2), 20.0% (1/5), and 48.9% (22/45), respectively. SNP array analysis in 138 cases revealed clinically significant submicroscopic aberrations in 4 karyotypically normal cases, with incremental detection rates of 2.0% (2/101) in the isolated group and 5.4% (2/37) in the non-isolated group (p > 0.05). The pregnancy termination rates were 25.5% (35/137) for the isolated group and 90.2% (46/51) for the non-isolated group. Among pregnancies with isolated OFCs and no clinically significant submicroscopic or microscopic chromosomal abnormalities, the termination rates were 11.8% (4/34) for CL, 3.8% (1/26) for CP, and 39.0% (30/77), respectively. Longitudinal follow-up of 46 cases with subsequent pregnancies revealed no recurrence of OFC. Conclusion Most fetal OFCs are isolated, presenting a very low risk of chromosomal abnormalities. For pregnancies with first-occurrence OFCs, the integration of karyotyping and SNP array analysis effectively evaluates the genetic etiology and provides essential guidance for prenatal counseling and future pregnancy management. orofacial clefts cleft lip cleft palate chromosomal abnormality isolated non-isolated Introduction Orofacial clefts (OFCs) are among the most common birth defects worldwide, characterized by a failure of fusion in the lip and/or palate during early embryonic development. Typical OFCs are categorized into cleft lip (CL), cleft palate (CP), and cleft lip and palate (CLP) [ 1 ] . According to a recent study by Heydari et al., the global prevalence of these conditions is 0.3, 0.33, and 0.45 per 1000 live births, respectively [ 2 ] . In China, the prevalence is higher, at approximately 1.4 per 1000 live births [ 3 ] . Ultrasonography is the preferred method for prenatal screening of fetal facial malformations. Ultrasonography remains the primary tool for prenatal screening of OFCs. While clefts can be detected as early as the first trimester [ 4 , 5 ] , most cases are diagnosed after 20 weeks of gestation. In China, 20 to 24 weeks of gestation is recommended to be the best period for ultrasound examination [ 6 ] . The etiology of OFCs is multifactorial, involving genetic, environmental, and their interaction [ 7 ] . Syndromic clefts, such as those associated with trisomy 13, DiGeorge syndrome, and Pierre Robin sequence, are frequently accompanied by systemic malformations [ 8 , 9 ] . Non-syndromic clefts, which account for approximately 70% of cases, often occur as isolated anomalies without additional structural abnormalities [ 10 ] [ 11 ] . Genetic contributors to OFCs include chromosomal abnormalities, submicroscopic copy number variations (CNVs), and multiple gene mutations. Prenatal diagnosis, incorporating detailed ultrasound scanning and genetic testing, plays a critical role in assessing prognosis and guiding pregnancy management. Following a prenatal diagnosis of OFCs, detailed ultrasound or MRI examinations should be conducted to detect any additional malformations. In cases where fetuses present with isolated OFCs and are confirmed to be free of genetic abnormalities, postnatal surgical repair typically yields favorable outcomes. In recent years, advances in surgical techniques, coupled with an increased focus on health-related quality of life among patients and their families [ 12 ] , have fostered more active participation in pregnancy-related decision-making. In this retrospective study, we analyzed 205 pregnancies with first-occurrence OFCs over a 14-year period, incorporating a systematic assessment of prenatal genetic evaluation data, imaging findings, and pregnancy outcomes. Materials and Methods Patients and samples A total of 206 pregnancies with first-occurrence OFCs were referred for prenatal genetic testing at the Medical Genetics Diagnosis and Treatment Center of Fujian Provincial Maternity and Child Health Hospital. One case was excluded due to both parents having OFCs, leaving 205 pregnancies for analysis. Gestational ages at diagnosis ranged from 16 to 34 weeks. The specimens included 142 amniotic fluid samples collected before 24 weeks of gestation and 63 cord blood samples collected after 24 weeks of gestation. Pregnancy outcomes and subsequent pregnancy data were obtained from hospital records and telephone interviews. Ethical approval was granted by the local Ethics Committee, and written informed consent was obtained from all participants. Methods The following genetic diagnosis methods have been described in our previous studies [ 13 , 14 ] . Conventional karyotyping, involving cell culture, and G-banded karyotyping, was performed on all 205 samples according to standard protocols in our laboratory. The karyotypes were determined at a resolution of 320 to 500 bands and described according to the International System for Human Cytogenetic Nomenclature (ISCN) 2020 guidelines [ 15 ] . Single nucleotide polymorphism array (SNP array) analysis was conducted on 138 cases in the cohort using the Affymetrix CytoScan 750K array (Affymetrix Inc., Santa Clara, CA, USA). The Chromosome Analysis Suite (CHAS) 3.2 software (Affymetrix, Santa Clara, CA, USA) was utilized for raw data analysis. CNVs were classified according to the American College of Medical Genetics (ACMG) [ 16 ] . Detailed experimental processes and reporting criteria was performed as previously described [ 13 ] . Parental SNP array analysis was recommended for determining the inheritance of CNVs. Chromosomal aneuploidies, unbalanced structural rearrangements, as well as pathogenic and likely pathogenic CNVs were designated as clinically significant aberrations. The data were analyzed using SPSS software v26.0 (SPSS Inc., Chicago, IL, USA). Statistical comparisons were performed using the chi-square test, the Fisher's exact test, and p < 0.05 was considered statistically significant. Results Baseline Characteristics The baseline characteristics of the cohort are detailed in Table 1 . The study included 42 cases of CL, 31 cases of CP, and 132 cases of CLP. Cases were classified as isolated (n = 153) or non-isolated (n = 52) OFCs based on the presence of additional structural malformations. The proportion of isolated cases was highest in CL (95.2%, 40/42), followed by CP (83.8%, 26/31) and CLP (65.9%, 87/132). Among the 52 non-isolated OFC cases, 34 were associated with multisystem malformations and 18 with single-system malformations. Table 1 The basic characteristics of the 205 cases with typical orofacial cleft. Characteristics Value Gestational weeks at invasive prenatal diagnosis (weeks) (mean ± SD) 24.2 ± 3.0 Maternal age (years) 29.0 ± 4.3 OFCs type Cleft lip (n, %) 42, 20.0% Cleft palate (n, %) 31,15.1% Cleft lip and palate (n, %) 132, 64.4% OFCs Classification Isolated (n, %) 152, 74.2% Non-isolated ( n, %) 53, 25.9% Specimens Amniotic fluid (n, %) 142, 69.3% Cord blood (n, %) 63, 30.7% OFCs, Orofacial clefts Chromosomal abnormalities detected by conventional karyotyping Chromosomal abnormalities were identified in 24 cases (11.7%) through conventional karyotyping. Trisomy 13 was the most frequent abnormality, accounting for 50.0% (12/24) of cases, followed by trisomy 18 at 20.8% (5/24). All chromosomal aberrations occurred in the non-isolated group, yielding a chromosomal abnormality rate of 46.2% (24/52) for non-isolated OFCs. Detailed results are summarized in Table 2 . Among non-isolated cases, the chromosomal abnormality rates for those with multisystem and single-system malformations were 47.1% (16/34) and 44.4% (8/18), respectively (p > 0.05). Stratified by cleft type, the chromosomal abnormality rates for non-isolated CL, CP, and CLP were 50.0% (1/2), 20.0% (1/5), and 48.9% (22/45), respectively. Table 2 Details of the 24 fetuses with abnormal karyotypes. Case Gestational age at invasive prenatal diagnosis (weeks) Karyotypes/Associated syndrome Ultrasound findings Outcomes 1 21 47,XY,+18 /Edward's syndrome CLP, FGR, microtia, VSD TOP 2 29 47,XX,+18 /Edward's syndrome CLP,omphalocele, VSD, Single umbilical artery, hyperechoic kidney, NF thickening, EIF TOP 3 24 47,XY,+18 /Edward's syndrome CLP, VSD, microtia, single umbilical artery TOP 4 29 47,XY,+12[4]/46,XY[42] CLP, thoraco-gastroschisis, cranial coloboma with meningoencephalocele, ectopia cordis, VSD, double outlet right ventricle, pulmonary artery stenosis, strephenopodia, abnormal blood flow spectrum of middle cerebral artery, umbilical artery TOP 5 24 47,XX,+18 /Edward's syndrome CLP, bilateral choroid plexus cysts, strawberry head, small jaw, microtia, VSD, mild mitral regurgitation, hyperechoic kidney, overlapping fingers TOP 6 23 47,XY,+13 /Patau's syndrome CLP, agenesis of the corpus callosum or lobate holoprosencephalon, omphalocele, VSD, bilateral kidney enlargement, NF thickening TOP 7 34 47,XY,+13 /Patau's syndrome CLP, VSD, aortic stenosis, absence of the inferior vermis of the cerebellum, posterior fossa pool broadening, EIF TOP 8 18 47,XY,+18/Edward's syndrome CLP, VSD, aortic stenosis TOP 9 24 46,XY,rob(13)(q10;q10) /Patau's syndrome CLP, holoprosencephaly, hypertelorism, flat nose, omphalocele TOP 10 24 47,XY,+13 /Patau's syndrome VSD, Tetralogy of Fallot, tricuspid regurgitation, hypertelorism, microphthalmia, omphalocele, left polycystic kidney, left ureteral dilatation, left ureterocele TOP 11 21 46,XY,der(22)(q10;q10) CLP, FGR, enlarged cardiothoracic ratio hypoplasia of the inferior cerebellar vermis oligohydramnios TOP 12 27 46,XY,der(13;14),+13 /Patau's syndrome CLP, holoprosencephaly, VSD, aortic stenosis, omphalocele, sacrococcygeal malformations TOP 13 28 46,XY,der(22)(q10;q10) CLP, FGR, abnormal blood flow in the middle cerebral artery, arachnoid cyst, left-sided microphthalmia, micrognathia, cervical lymphatic cystic hygroma,situs inversus cardiac abnormalities (dextrocardia, pulmonary artery stenosis), low conus medullaris TOP 14 31 47,XY,+13 /Patau's syndrome CL, holoprosencephaly, EIF TOP 15 21 47,XY,+13 /Patau's syndrome CLP, omphalocele, polydactyly and overlapping fingers on the right hand, NF thickening Hypoplasia of the cerebellar vermis,Hypoplastic left heart, aortic stenosis, EIF TOP 16 29 47,XX,+13 /Patau's syndrome CLP, ventriculomegaly, cerebellar hypoplasia, absence of the inferior cerebellar vermis, microphthalmia, TOP 17 23 47,XY,+13 /Patau's syndrome CL, enlarged right ventricle, EIF TOP 18 24 46,XX,der(4)t(2;4)(p11;p11)dn CLP, FGR, VSD, hypoplastic left heart, mitral atresia, aortic atresia, cervical and dorsal lymphatic cystic hygroma, small kidneys TOP 19 30 47,XX,+13 /Patau's syndrome CLP, holoprosencephaly, complex cardiac malformations TOP 20 27 47,XX,+13 /Patau's syndrome CLP, VSD, holoprosencephaly, hypoplastic left heart, double outlet right ventricle, pulmonary artery stenosis, possible duplex right kidney, single umbilical artery TOP 21 30 46,XX,del(8)(q21.1q22.2) CP, NT thinckening, VSD, nasal bone dysplasia TOP 22 38 47,XX,+i(12)(p10)[7]/46,XX[104] CP, thickened skin on the nasal tip, NF thickening, TOP 23 24 47,XY,+13 /Patau's syndrome CLP, cerebellar malformation, bilateral microphthalmia, microtia, VSD, right subclavian artery anomaly, polydactyly in both hands, overlapping fingers, polydactyly in both feet, low conus medullaris, dysplastic sacral vertebrae TOP 24 24 47,XXX LP, left choroid plexus cyst, conus medullaris at L4 LB CL, Cleft lip; CLP, Cleft lip and palate; CP, Cleft Palate; NT, nuchal translucency; TOP, termination of pregnancy; VSD, ventricular septal defects; FGR, fetal growth restriction; EIF, echogenic intracardiac focus; TOP: termination of pregnancy; LB, live birth Submicroscopic aberrations detected by SNP array analysis Among the 138 cases that underwent both karyotyping and SNP array analysis, submicroscopic chromosomal aberrations were identified in 12 cases, including 4 cases with clinically significant submicroscopic abnormalities in individuals with normal karyotypes. The detection rates of additional clinically significant submicroscopic abnormalities were 2.0% (2/101) in the isolated group and 5.4% (2/37) in the non-isolated group (p > 0.05). Detailed results are presented in Table 3 . Table 3 Submicroscopic chromosomal aberrations detected by SNP array analysis. Case number Ultrasound findings SNP array analysis Parental inheritance Clinical significance Pregnancy outcome 25 CL, left echogenic intracardiac focus arr[GRCh37] 22q11.22q11.23(22997929–25041592)×3 NA VOUS LB 26 CLP arr[GRCh37] 22q12.3(33,854,040 − 34,400,183)×3 NA VOUS NA 27 CP arr[GRCh37] 8q24.22q24.3(134,714,740 − 146,292,734)×2 hmz / VOUS LB 28 CLP, craniosynostoses arr[GRCh37] Xp22.33(288,066 − 1,023,657)×1 dn Pathogenic TOP 29 CP, tricuspid regurgitation arr[GRCh37] 5q23.3q31.1(129,930550-131,441,100)×3 Maternal Likely benign LB 30 CP arr[GRCh37] 15q11.2q13.1(23693931 28526905)×3 dn Pathogenic TOP 31 CLP, Ventricular septal defects arr[GRCh37] 4q35.1q35.2(186031030–189191908)×3 Paternal VOUS LB 32 CLP arr[GRCh37] 15q22.33q25.1(67432515–78796221)×2 hmz / VOUS TOP 33 CLP, VSD, thymusv dysplasia arr[GRCh37] 22q11.21(18916843–21464764)×1 dn Pathogenic TOP 34 CLP arr[GRCh37] 6p25.3p25.2(993944_2513338)×3 Paternal VOUS LB 35 CP arr[GRCh37] 7q11.23(72669481_74154209)×1 Maternal Pathogenic LB 36 CLP arr[GRCh37] 5q21.3(106340917–107886317)×1 / VOUS TOP CL, Cleft lip; CLP, Cleft lip and palate; CP, Cleft Palate; NA, not available; VOUS, variants of uncertain significance; TOP, termination of pregnancy; VSD, ventricular septal defects; dn, De novo, LB, live birth Pregnancy outcome Pregnancy outcome data were available for 188 cases (91.7%). Of the 24 pregnancies with clinically significant aberrations, all were terminated except for case 35 (Table 3 ). This fetus presented with cleft palate and a 1.48 Mb deletion at 7q11.23, inherited from a mother with intellectual disability. Among the remaining 160 cases, the termination rate was significantly higher in the non-isolated group at 80.8% (21/26) compared to 23.0% (31/135) in the isolated OFC group (p < 0.05). Within the isolated group without clinically significant aberrations, the termination rate for CLP was 39.0% (30/77), which was significantly higher (p < 0.05) than that for CL at 11.8% (4/34) and for CP at 3.8% (1/26). Follow-up data on subsequent pregnancies were obtained for 45 cases, and no recurrent OFC were observed. Discussion The distribution of different types of OFCs varied across studies [ 2 , 3 , 17 ] . In our study, CLP was more frequently observed than CL or CP. In our study, CLP was more frequently observed than CL or CP. OFCs can occur either in isolation or in combination with other system malformations. Notably, over two-thirds of the cases in this study were isolated OFCs. Since CMA was not widely applied in prenatal diagnosis at our center before 2016, a portion of fetuses with OFCs underwent only conventional karyotyping. In isolated OFCs, no chromosomal abnormalities were detected by conventional karyotyping. By contrast, the chromosomal abnormality rate was as high as 46.2% in the non-isolated group. Consistent with previous studies [ 18 , 19 ] , there was no significant difference in the chromosomal abnormality rates between cases with single-system malformations and those with multisystem malformations. This finding emphasizes that the risk of chromosomal abnormalities is considerably higher when OFCs are accompanied by other structural malformations. Trisomy 13 has been reported as the most frequent chromosomal abnormality in OFC cases, followed by trisomy 18 [ 20 – 23 ] . Consistent with these findings, our study showed that trisomy 13 accounted for 50.0% of karyotype-detectable abnormalities, followed by trisomy 18 (5 cases, 21.7%) and trisomy 22 (2 cases, 8.7%). This underscores the diagnostic utility of conventional karyotyping in identifying genetic etiologies when OFCs are associated with additional major malformations. Additionally, two cases of low-level mosaicism involving chromosome 12 were detected in this study. Mosaic trisomy 12 is a rare condition, with few prenatal and postnatal cases reported in the literature. Previous studies have highlighted a spectrum of ultrasound findings, from normal to severe congenital anomalies, in pregnancies with trisomy 12 mosaicism [ 24 – 27 ] . One of our cases exhibited 8.9% (4/45) mosaicism in cultured amniocytes and presented with CLP accompanied by multisystem defects involving cardiovascular, abdominal, cranial, and skeletal development. The discordance between low-level mosaicism and severe ultrasound abnormalities might result from the differential distribution of abnormal cells in various tissues, although further testing was unavailable to confirm this hypothesis. Another case involved mosaicism with an extra isochromosome 12p (i(12p)), responsible for Pallister-Killian syndrome (PKS, OMIM: #601803). This case has been detailed in a previous publication [ 28 ] . In recent years, chromosomal microarray analysis (CMA) has confirmed the significant role of CNVs in the genetic etiology of OFCs [ 29 – 32 ] . In this study, SNP array analysis identified four pathogenic/likely pathogenic CNVs (2.9%), a detection rate notably lower than postnatal reports [ 32 , 33 ] . The 22q11.2 deletion syndrome, associated with DiGeorge syndrome (OMIM: #188400) and velocardiofacial syndrome (OMIM: #192430), is the most common syndrome linked to conotruncal anomalies [ 34 , 35 ] . Mechanistically, haploinsufficiency of TBX1 , encoding a T-box transcription factor essential for pharyngeal arch development, disrupts cranial neural crest cell migration, leading to defective palatal shelf elevation and thymic hypoplasia. A study by Bashir et al. recommended routine 22q11 fluorescence in situ hybridization (FISH) testing for children with CL and/or CP due to the diagnostic implications of velocardiofacial syndrome [ 36 ] . In present study, we identified a 2.5 Mb microdeletion at 22q11.2 in a fetus with CP, heart defects and thymus gland dysplasia, provides critical prenatal validation of this association. Additionally, a de novo 4.8 Mb at 15q11.2-q13.1 was detected in a fetus with isolated CP. This region encompasses the critical region of 15q11-q13 duplication syndrome (OMIM: #608636), which is typically characterized by neurodevelopmental disorders (autism, epilepsy) and hypotonia [ 37 ] . To the best of our knowledge, this duplication has not been previously reported in cases of OFCs, which expands the phenotypic spectrum of 15q11-q13 CNVs and highlights potential genotype-phenotype correlations requiring further validation. In a male fetus with CLPand craniosynostoses, a 736 Kb deletion was identified at Xp22.33, involving the Short Stature Homeobox (SHOX) gene. Haploinsufficiency of SHOX is primarily associated with Léri-Weill dyschondrosteosis [ 38 , 39 ] and facial dysmorphism has been observed in some cases with Xp22.33 microdeletions [ 40 , 41 ] . A study by Chen et al. concluded that SHOX2 plays a critical role in osteogenesis of a specific cell lineage and contributes to the development of the palatine process of the maxilla by interacting with distal cis-regulatory elements to regulate skeletogenic gene expression and pattern the hard palate [ 42 ] . Therefore, the relationship between OFCs and Shox is worthy of further in-depth study. Furthermore, a 1.5 Mb deletion at 7q11.23, responsible for Williams-Beuren syndrome (WBS; OMIM: #194050), was identified in a fetus with isolated CP. The most common manifestations of WBS include a characteristic craniofacial morphology, developmental delays, cognitive and behavioural changes [ 43 ] . Reports of WBS patients with cleft palate are not uncommon [ 44 – 47 ] , suggesting partial phenotypic overlap with this congenital anomaly. Genetic analysis confirmed maternal inheritance of the deletion. While the mother’s primary manifestation was intellectual disability, the fetus developed CP and may face heightened risks of multisystem involvement due to allelic dosage effects or variable expressivity of the 7q11.23 deletion. Following comprehensive genetic counseling regarding prognosis and recurrence risks, the family opted to continue the pregnancy. Regarding pregnancy outcomes, genetic abnormalities of clinical significance and non-isolated OFCs were the primary reasons for pregnancy termination. Additionally, we placed special attention to isolated OFCs. CLP cases had the highest termination rate, whereas all 22 cases of isolated CP were continued. Notably, follow-up of 45 cases revealed no recurrence of OFCs in subsequent pregnancies. These findings suggest that karyotyping and SNP array analysis are effective in identifying the genetic etiologies of primary OFCs, thereby providing valuable guidance for future pregnancies. This study has several limitations. Its retrospective design and the fact that not all cases underwent SNP array analysis may have introduced bias in evaluating the incidence of CNVs and single-gene disorders in fetuses with OFCs. In conclusion, this study underscores the effectiveness of comprehensive genetic evaluations, including karyotyping and SNP array analysis, in pregnancies with first-occurrence OFCs. Notably, chromosomal karyotype abnormalities are unlikely to cause isolated OFCs. Moreover, pregnancy outcomes for OFCs are primarily determined by the chromosomal analysis results, the specific type of OFCs, and the presence of other systemic malformation. Abbreviations OFCs Orofacial clefts CL cleft lip CP cleft palate CLP cleft lip and palate CNVs copy number variations ISCN International System for Human Cytogenetic Nomenclature CHAS Chromosome Analysis Suite ACMG American College of Medical Genetics CMA chromosomal microarray analysis FISH fluorescence in situ hybridization SHOX Short Stature Homeobox WBS Williams-Beuren syndrome Declarations Ethics approval and consent to participate The present study was approved by the Protection of Human Ethics Committee of Fujian Maternity and Child Health Hospital. Written informed consent was obtained from each pregnant woman. Consent for publication Written informed consent to publish this case was obtained from all the participants and parents of the minor participants, including case description and medical data. Availability of data and material Not applicable Competing interests The authors declare they have no conflict of interest. Funding The study was supported by Key Special Projects of Fujian Provincial Department of Science and Technology (grant No. 2021YZ034011). Innovation Platform Project of Science and Technology, Fujian province (2021Y2012) National Key Clinical Specialty Construction Program of China (Obstetric). Authors' contributions XW and XX drafted the main manuscript. JL, LS and BL prepared Tables 1–3, NL and LX provided genetic counseling. DG, LZreviewed and revised the manuscript. All authors reviewed the manuscript. Acknowledgements Not applicable References Vyas T, P Gupta, S Kumar, R Gupta, T Gupta, and H P Singh, Cleft of lip and palate: A review. J Family Med Prim Care, 2020; 9(6): p. 2621-2625. Salari N, N Darvishi, M Heydari, S Bokaee, F Darvishi, and M Mohammadi, Global prevalence of cleft palate, cleft lip and cleft palate and lip: A comprehensive systematic review and meta-analysis. J Stomatol Oral Maxillofac Surg, 2022; 123(2): p. 110-120. Wang M, Y Yuan, Z Wang, D Liu, Z Wang, F Sun, et al., Prevalence of Orofacial Clefts among Live Births in China: A Systematic Review and Meta-Analysis. Birth Defects Res, 2017; 109(13): p. 1011-1019. Tonni G, G Grisolia, and W Sepulveda, Early prenatal diagnosis of orofacial clefts: evaluation of the retronasal triangle using a new three-dimensional reslicing technique. Fetal Diagn Ther, 2013; 34(1): p. 31-7. Pekar-Zlotin M, N Zilberman Sharon, Y Melcer, Y Tal-Bliman, J Ezratty, M Feingold-Zadok, et al., Pregnancy with Facial Cleft: 20 Years of Experience at a Single Center. Isr Med Assoc J, 2023; 25(10): p. 678-682. Expert Group of Multidisciplinary Treatment Process of Cleft L and Palate, [Specification of the multidisciplinary treatment process of cleft lip and palate during pregnancy, prenatal and postnatal stages]. Zhonghua Kou Qiang Yi Xue Za Zhi, 2021; 56(11): p. 1059-1065. Beaty T H, M L Marazita, and E J Leslie, Genetic factors influencing risk to orofacial clefts: today's challenges and tomorrow's opportunities. F1000Res, 2016; 5: p. 2800. Babai A and M Irving, Orofacial Clefts: Genetics of Cleft Lip and Palate. Genes (Basel), 2023; 14(8). Schindewolf E, N Khalek, M P Johnson, J Gebb, B Coleman, T B Crowley, et al., Expanding the fetal phenotype: Prenatal sonographic findings and perinatal outcomes in a cohort of patients with a confirmed 22q11.2 deletion syndrome. Am J Med Genet A, 2018; 176(8): p. 1735-1741. Li S, A Chao, Z Li, C A Moore, Y Liu, J Zhu, et al., Folic acid use and nonsyndromic orofacial clefts in China: a prospective cohort study. Epidemiology, 2012; 23(3): p. 423-32. Salazar Trujillo A, C Rincón-Guio, L López Narváez, J Cáceres, and J D Charry, First trimester sonographic diagnosis of orofacial defects. Review of literature. J Matern Fetal Neonatal Med, 2020; 33(18): p. 3200-3206. Fan K L, C K Black, E Mantilla-Rivas, D I Bulas, E Rubio, A R Blask, et al., Coordination of the Fetal Medicine Institute and the Cleft and Craniofacial Center: Application to Early Management of Infants With Cleft Lip and Palate. J Craniofac Surg, 2019; 30(7): p. 2061-2064. Wu X, Y Li, L Su, X Xie, M Cai, N Lin, et al., Chromosomal Microarray Analysis for the Fetuses with Aortic Arch Abnormalities and Normal Karyotype. Mol Diagn Ther, 2020; 24(5): p. 611-619. Wu X, L Xu, Y Li, N Lin, L Su, M Cai, et al., Submicroscopic aberrations of chromosome 16 in prenatal diagnosis. Mol Cytogenet, 2019; 12: p. 36. Mcgowan-Jordan J, R J Hastings, and S Moore, An International System for Human Cytogenomic Nomenclature (2020). Cytogenetic and genome research, 2020. South S T, C Lee, A N Lamb, A W Higgins, H M Kearney, G Working Group for the American College of Medical, et al., ACMG Standards and Guidelines for constitutional cytogenomic microarray analysis, including postnatal and prenatal applications: revision 2013. Genet Med, 2013; 15(11): p. 901-9. Lee C W, S M Hwang, Y S Lee, M A Kim, and K Seo, Prevalence of orofacial clefts in Korean live births. Obstet Gynecol Sci, 2015; 58(3): p. 196-202. Beriaghi S, S Myers, S Jensen, S Kaimal, C Chan, and G B Schaefer, Cleft lip and palate: association with other congenital malformations. Journal of Clinical Pediatric Dentistry, 2009; 33(3): p. 207-210. Azadgoli B, N C O Munabi, A Fahradyan, A Auslander, M McCullough, N Aflatooni, et al., Congenital Heart Disease in Patients With Cleft Lip/Palate and Its Impact on Cleft Management. Cleft Palate Craniofac J, 2020; 57(8): p. 957-966. Petry P, J B Polli, V F Mattos, R C Rosa, P R Zen, C Graziadio, et al., Clinical features and prognosis of a sample of patients with trisomy 13 (Patau syndrome) from Brazil. Am J Med Genet A, 2013; 161a(6): p. 1278-83. Bergé S J, H Plath, P T Van de Vondel, T Appel, B Niederhagen, J J Von Lindern, et al., Fetal cleft lip and palate: sonographic diagnosis, chromosomal abnormalities, associated anomalies and postnatal outcome in 70 fetuses. Ultrasound Obstet Gynecol, 2001; 18(5): p. 422-31. Bruns D, Birth history, physical characteristics, and medical conditions in long-term survivors with full trisomy 13. Am J Med Genet A, 2011; 155a(11): p. 2634-40. Vibert F, G Schmidt, K Löffler, A Gasiorek-Wiens, W Henrich, and S Verlohren, Accuracy of prenatal detection of facial clefts and relation between facial clefts, additional malformations and chromosomal abnormalities: a large referral-center cohort. Arch Gynecol Obstet, 2024; 309(5): p. 1971-1980. Chen C P, Y N Su, J W Su, S R Chern, Y T Chen, L F Chen, et al., Mosaic trisomy 12 at amniocentesis: prenatal diagnosis and molecular genetic analysis. Taiwan J Obstet Gynecol, 2013; 52(1): p. 97-105. Chen C P, C J Lin, S R Chern, P S Wu, Y N Chen, S W Chen, et al., Prenatal diagnosis and molecular cytogenetic characterization of low-level mosaic trisomy 12 at amniocentesis associated with a favorable pregnancy outcome. Taiwan J Obstet Gynecol, 2017; 56(2): p. 238-242. Roberts W, A Zurada, A Zurada-ZieliŃSka, J Gielecki, and M Loukas, Anatomy of trisomy 12. Clin Anat, 2016; 29(5): p. 633-7. Hong B, J Zunich, A Openshaw, and R M Toydemir, Clinical features of trisomy 12 mosaicism-Report and review. Am J Med Genet A, 2017; 173(6): p. 1681-1686. Wu X, X Xie, L Su, N Lin, B Liang, N Guo, et al., Prenatal diagnosis of Pallister-Killian syndrome and literature review. J Cell Mol Med, 2021; 25(18): p. 8929-8935. Lansdon L A, B W Darbro, A L Petrin, A M Hulstrand, J M Standley, R B Brouillette, et al., Identification of Isthmin 1 as a Novel Clefting and Craniofacial Patterning Gene in Humans. Genetics, 2018; 208(1): p. 283-296. Cai Y, K E Patterson, F Reinier, S E Keesecker, E Blue, M Bamshad, et al., Copy Number Changes Identified Using Whole Exome Sequencing in Nonsyndromic Cleft Lip and Palate in a Honduran Population. Birth Defects Res, 2017; 109(16): p. 1257-1267. Conte F, M Oti, J Dixon, C E Carels, M Rubini, and H Zhou, Systematic analysis of copy number variants of a large cohort of orofacial cleft patients identifies candidate genes for orofacial clefts. Hum Genet, 2016; 135(1): p. 41-59. Lei T Y, H T Wang, F Li, Y Q Cui, F Fu, R Li, et al., Application of high resolution SNP arrays in patients with congenital oral clefts in south China. J Genet, 2016; 95(4): p. 801-809. da Silva H P V, G H M Oliveira, M A G Ururahy, J F Bezerra, K S C de Souza, R H Bortolin, et al., Application of high-resolution array platform for genome-wide copy number variation analysis in patients with nonsyndromic cleft lip and palate. J Clin Lab Anal, 2018; 32(6): p. e22428. McDonald-McGinn D M and K E Sullivan, Chromosome 22q11.2 deletion syndrome (DiGeorge syndrome/velocardiofacial syndrome). Medicine (Baltimore), 2011; 90(1): p. 1-18. Lewyllie A, J Roosenboom, K Indencleef, P Claes, A Swillen, K Devriendt, et al., A Comprehensive Craniofacial Study of 22q11.2 Deletion Syndrome. J Dent Res, 2017; 96(12): p. 1386-1391. Bashir M A, P D Hodgkinson, T Montgomery, and M Splitt, 22q11 Deletion in children with cleft lip and palate--is routine screening justified? J Plast Reconstr Aesthet Surg, 2008; 61(2): p. 130-2. Urraca N, J Cleary, V Brewer, E K Pivnick, K McVicar, R L Thibert, et al., The interstitial duplication 15q11.2-q13 syndrome includes autism, mild facial anomalies and a characteristic EEG signature. Autism Res, 2013; 6(4): p. 268-79. Shima H, T Tanaka, T Kamimaki, S Dateki, K Muroya, R Horikawa, et al., Systematic molecular analyses of SHOX in Japanese patients with idiopathic short stature and Leri-Weill dyschondrosteosis. J Hum Genet, 2016; 61(7): p. 585-91. Vannelli S, M Baffico, R Buganza, F Verna, G Vinci, D Tessaris, et al., SHOX deficiency in children with growth impairment: evaluation of known and new auxological and radiological indicators. Ital J Pediatr, 2020; 46(1): p. 163. Depeyre A, M Schlund, R Nicot, and J Ferri, Dental and Maxillofacial Signs in Leri-Weill Dyschondrosteosis. J Oral Maxillofac Surg, 2019; 77(4): p. 762-768. Dupont C, A Lebbar, C Teinturier, F Baverel, G Viot, D Le Tessier, et al., First reported case of intrachromosomal cryptic inv dup del Xp in a boy with developmental retardation. Am J Med Genet A, 2007; 143a(11): p. 1236-43. Xu J, L Wang, H Li, T Yang, Y Zhang, T Hu, et al., Shox2 regulates osteogenic differentiation and pattern formation during hard palate development in mice. J Biol Chem, 2019; 294(48): p. 18294-18305. Kozel B A, B Barak, C A Kim, C B Mervis, L R Osborne, M Porter, et al., Williams syndrome. Nat Rev Dis Primers, 2021; 7(1): p. 42. Blanco-Dávila F and J A Olveda-Rodriguez, Cleft palate in a patient with Williams' syndrome. J Craniofac Surg, 2001; 12(2): p. 145-7. Domenico S, C Orlando, F F Graziana, P Papi, and A Giulia, Cleft palate in Williams syndrome. Ann Maxillofac Surg, 2013; 3(1): p. 84-6. Vincent C, J M Mercier, and A David, [Cleft palate and Williams syndrome]. Rev Stomatol Chir Maxillofac, 2008; 109(1): p. 44-7. Yamaguchi T, T Shirota, M Adel, M Takahashi, S Haga, R Nagahama, et al., Orthodontic Treatment and Maxillary Anterior Segmental Distraction Osteogenesis of a Subject with Williams-Beuren Syndrome and Isolated Cleft Palate: A Long-Term Follow-Up from the Age of 5 to 24 Years. Case Rep Dent, 2017; 2017: p. 7019045. Additional Declarations No competing interests reported. Cite Share Download PDF Status: Posted Version 1 posted 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-6191104","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":434229940,"identity":"a8000f90-958b-4c6d-83bd-c3b972add8ce","order_by":0,"name":"Xiaoqing Wu","email":"","orcid":"","institution":"Fujian Women and Children Hospital","correspondingAuthor":false,"prefix":"","firstName":"Xiaoqing","middleName":"","lastName":"Wu","suffix":""},{"id":434229941,"identity":"1722e825-e5c4-481b-82b2-cf40b7638331","order_by":1,"name":"Xiaorui Xie","email":"","orcid":"","institution":"Fujian Women and Children Hospital","correspondingAuthor":false,"prefix":"","firstName":"Xiaorui","middleName":"","lastName":"Xie","suffix":""},{"id":434229942,"identity":"93376077-6ccb-4463-ab9e-d2ff371cd6c4","order_by":2,"name":"Jinzhou Lu","email":"","orcid":"","institution":"Fujian Medical University","correspondingAuthor":false,"prefix":"","firstName":"Jinzhou","middleName":"","lastName":"Lu","suffix":""},{"id":434229943,"identity":"e7de7daf-c3b6-4b68-b97b-0baf7d4fe6df","order_by":3,"name":"Linjuan Su","email":"","orcid":"","institution":"Fujian Women and Children Hospital","correspondingAuthor":false,"prefix":"","firstName":"Linjuan","middleName":"","lastName":"Su","suffix":""},{"id":434229944,"identity":"b67600b1-66c2-4031-a1d1-aeda181fa823","order_by":4,"name":"Bin Liang","email":"","orcid":"","institution":"Fujian Women and Children Hospital","correspondingAuthor":false,"prefix":"","firstName":"Bin","middleName":"","lastName":"Liang","suffix":""},{"id":434229945,"identity":"02918300-bfa9-4e27-9a29-c6ac2eefc1e0","order_by":5,"name":"Na Lin","email":"","orcid":"","institution":"Fujian Women and Children Hospital","correspondingAuthor":false,"prefix":"","firstName":"Na","middleName":"","lastName":"Lin","suffix":""},{"id":434229946,"identity":"daaf2e6d-b322-4780-a4c1-79b0da44801d","order_by":6,"name":"Liangpu Xu","email":"","orcid":"","institution":"Fujian Women and Children Hospital","correspondingAuthor":false,"prefix":"","firstName":"Liangpu","middleName":"","lastName":"Xu","suffix":""},{"id":434229947,"identity":"931cff84-d994-422a-9f10-352a104c5df6","order_by":7,"name":"Danhua Guo","email":"","orcid":"","institution":"Fujian Women and Children Hospital","correspondingAuthor":false,"prefix":"","firstName":"Danhua","middleName":"","lastName":"Guo","suffix":""},{"id":434229948,"identity":"bc4704f1-01fb-4030-84ca-89c5375d8709","order_by":8,"name":"Lin Zheng","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAAwklEQVRIiWNgGAWjYDACdsYGhsQ/NXL8zMwHHxCnhRmo5WHDMWPJdrZkAyK1MDAwPmxgTjQ4z2MmQJQO+WbmtgeJO9gSjA8zmDEw1NhEE9TC2MzYbpB4RibP7DBD2gOGY2m5DQTdxczYJpHAxlYM1HLcgLHhMGEtbBAtzImbm4EMorTwgLQktjEnbmBmZiNOiwTYljPHjCUOszEbJBDjF/n29meSPyqAUdl//uODDzU2hLWgggTSlI+CUTAKRsEowAUATNU5ayOKOdQAAAAASUVORK5CYII=","orcid":"","institution":"Fujian Women and Children Hospital","correspondingAuthor":true,"prefix":"","firstName":"Lin","middleName":"","lastName":"Zheng","suffix":""}],"badges":[],"createdAt":"2025-03-10 02:23:13","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-6191104/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-6191104/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":80630425,"identity":"c62ee19b-8f48-4b6c-a874-83743a9853fb","added_by":"auto","created_at":"2025-04-15 11:38:47","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":901401,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-6191104/v1/fdd82eac-81c2-48fd-9562-5f4bd1988619.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Prenatal genetic evaluation and outcomes in pregnancies with first-occurrence typical orofacial clefts","fulltext":[{"header":"Introduction","content":"\u003cp\u003eOrofacial clefts (OFCs) are among the most common birth defects worldwide, characterized by a failure of fusion in the lip and/or palate during early embryonic development. Typical OFCs are categorized into cleft lip (CL), cleft palate (CP), and cleft lip and palate (CLP) \u003csup\u003e[\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e]\u003c/sup\u003e. According to a recent study by Heydari et al., the global prevalence of these conditions is 0.3, 0.33, and 0.45 per 1000 live births, respectively \u003csup\u003e[\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e]\u003c/sup\u003e. In China, the prevalence is higher, at approximately 1.4 per 1000 live births \u003csup\u003e[\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e]\u003c/sup\u003e. Ultrasonography is the preferred method for prenatal screening of fetal facial malformations. Ultrasonography remains the primary tool for prenatal screening of OFCs. While clefts can be detected as early as the first trimester \u003csup\u003e[\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e, \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e]\u003c/sup\u003e, most cases are diagnosed after 20 weeks of gestation. In China, 20 to 24 weeks of gestation is recommended to be the best period for ultrasound examination \u003csup\u003e[\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e]\u003c/sup\u003e.\u003c/p\u003e \u003cp\u003eThe etiology of OFCs is multifactorial, involving genetic, environmental, and their interaction \u003csup\u003e[\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e]\u003c/sup\u003e. Syndromic clefts, such as those associated with trisomy 13, DiGeorge syndrome, and Pierre Robin sequence, are frequently accompanied by systemic malformations \u003csup\u003e[\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e, \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e]\u003c/sup\u003e. Non-syndromic clefts, which account for approximately 70% of cases, often occur as isolated anomalies without additional structural abnormalities \u003csup\u003e[\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e] [\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e]\u003c/sup\u003e. Genetic contributors to OFCs include chromosomal abnormalities, submicroscopic copy number variations (CNVs), and multiple gene mutations. Prenatal diagnosis, incorporating detailed ultrasound scanning and genetic testing, plays a critical role in assessing prognosis and guiding pregnancy management.\u003c/p\u003e \u003cp\u003eFollowing a prenatal diagnosis of OFCs, detailed ultrasound or MRI examinations should be conducted to detect any additional malformations. In cases where fetuses present with isolated OFCs and are confirmed to be free of genetic abnormalities, postnatal surgical repair typically yields favorable outcomes. In recent years, advances in surgical techniques, coupled with an increased focus on health-related quality of life among patients and their families\u003csup\u003e[\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e]\u003c/sup\u003e, have fostered more active participation in pregnancy-related decision-making.\u003c/p\u003e \u003cp\u003eIn this retrospective study, we analyzed 205 pregnancies with first-occurrence OFCs over a 14-year period, incorporating a systematic assessment of prenatal genetic evaluation data, imaging findings, and pregnancy outcomes.\u003c/p\u003e"},{"header":"Materials and Methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003ePatients and samples\u003c/h2\u003e \u003cp\u003eA total of 206 pregnancies with first-occurrence OFCs were referred for prenatal genetic testing at the Medical Genetics Diagnosis and Treatment Center of Fujian Provincial Maternity and Child Health Hospital. One case was excluded due to both parents having OFCs, leaving 205 pregnancies for analysis. Gestational ages at diagnosis ranged from 16 to 34 weeks. The specimens included 142 amniotic fluid samples collected before 24 weeks of gestation and 63 cord blood samples collected after 24 weeks of gestation. Pregnancy outcomes and subsequent pregnancy data were obtained from hospital records and telephone interviews. Ethical approval was granted by the local Ethics Committee, and written informed consent was obtained from all participants.\u003c/p\u003e \u003c/div\u003e\n\u003ch3\u003eMethods\u003c/h3\u003e\n\u003cp\u003eThe following genetic diagnosis methods have been described in our previous studies \u003csup\u003e[\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e, \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e]\u003c/sup\u003e.\u003c/p\u003e \u003cp\u003eConventional karyotyping, involving cell culture, and G-banded karyotyping, was performed on all 205 samples according to standard protocols in our laboratory. The karyotypes were determined at a resolution of 320 to 500 bands and described according to the International System for Human Cytogenetic Nomenclature (ISCN) 2020 guidelines\u003csup\u003e[\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e]\u003c/sup\u003e.\u003c/p\u003e \u003cp\u003eSingle nucleotide polymorphism array (SNP array) analysis was conducted on 138 cases in the cohort using the Affymetrix CytoScan 750K array (Affymetrix Inc., Santa Clara, CA, USA). The Chromosome Analysis Suite (CHAS) 3.2 software (Affymetrix, Santa Clara, CA, USA) was utilized for raw data analysis. CNVs were classified according to the American College of Medical Genetics (ACMG) \u003csup\u003e[\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e]\u003c/sup\u003e. Detailed experimental processes and reporting criteria was performed as previously described \u003csup\u003e[\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e]\u003c/sup\u003e. Parental SNP array analysis was recommended for determining the inheritance of CNVs.\u003c/p\u003e \u003cp\u003eChromosomal aneuploidies, unbalanced structural rearrangements, as well as pathogenic and likely pathogenic CNVs were designated as clinically significant aberrations.\u003c/p\u003e \u003cp\u003eThe data were analyzed using SPSS software v26.0 (SPSS Inc., Chicago, IL, USA). Statistical comparisons were performed using the chi-square test, the Fisher's exact test, and p\u0026thinsp;\u0026lt;\u0026thinsp;0.05 was considered statistically significant.\u003c/p\u003e"},{"header":"Results","content":"\u003cdiv id=\"Sec6\" class=\"Section2\"\u003e \u003ch2\u003eBaseline Characteristics\u003c/h2\u003e \u003cp\u003eThe baseline characteristics of the cohort are detailed in Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e. The study included 42 cases of CL, 31 cases of CP, and 132 cases of CLP. Cases were classified as isolated (n\u0026thinsp;=\u0026thinsp;153) or non-isolated (n\u0026thinsp;=\u0026thinsp;52) OFCs based on the presence of additional structural malformations. The proportion of isolated cases was highest in CL (95.2%, 40/42), followed by CP (83.8%, 26/31) and CLP (65.9%, 87/132). Among the 52 non-isolated OFC cases, 34 were associated with multisystem malformations and 18 with single-system malformations.\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\u003eThe basic characteristics of the 205 cases with typical orofacial cleft.\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=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCharacteristics\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eValue\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGestational weeks at invasive prenatal diagnosis (weeks) (mean\u0026thinsp;\u0026plusmn;\u0026thinsp;SD)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e24.2\u0026thinsp;\u0026plusmn;\u0026thinsp;3.0\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMaternal age (years)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e29.0\u0026thinsp;\u0026plusmn;\u0026thinsp;4.3\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOFCs type\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCleft lip (n, %)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e42, 20.0%\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCleft palate (n, %)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e31,15.1%\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCleft lip and palate (n, %)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e132, 64.4%\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c2\" namest=\"c1\"\u003e \u003cp\u003eOFCs Classification\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eIsolated (n, %)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e152, 74.2%\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNon-isolated ( n, %)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e53, 25.9%\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSpecimens\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAmniotic fluid (n, %)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e142, 69.3%\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCord blood (n, %)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e63, 30.7%\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003ctfoot\u003e \u003ctr\u003e\u003ctd colspan=\"2\"\u003eOFCs, Orofacial clefts\u003c/td\u003e\u003c/tr\u003e \u003c/tfoot\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003c/div\u003e\n\u003ch3\u003eChromosomal abnormalities detected by conventional karyotyping\u003c/h3\u003e\n\u003cp\u003eChromosomal abnormalities were identified in 24 cases (11.7%) through conventional karyotyping. Trisomy 13 was the most frequent abnormality, accounting for 50.0% (12/24) of cases, followed by trisomy 18 at 20.8% (5/24). All chromosomal aberrations occurred in the non-isolated group, yielding a chromosomal abnormality rate of 46.2% (24/52) for non-isolated OFCs. Detailed results are summarized in Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e. Among non-isolated cases, the chromosomal abnormality rates for those with multisystem and single-system malformations were 47.1% (16/34) and 44.4% (8/18), respectively (p\u0026thinsp;\u0026gt;\u0026thinsp;0.05). Stratified by cleft type, the chromosomal abnormality rates for non-isolated CL, CP, and CLP were 50.0% (1/2), 20.0% (1/5), and 48.9% (22/45), respectively.\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\u003eDetails of the 24 fetuses with abnormal karyotypes.\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=\"char\" char=\".\" 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\u003eCase\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eGestational age at invasive prenatal diagnosis (weeks)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eKaryotypes/Associated syndrome\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eUltrasound findings\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eOutcomes\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=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e21\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e47,XY,+18 /Edward's syndrome\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eCLP, FGR, microtia, VSD\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eTOP\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=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e29\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e47,XX,+18 /Edward's syndrome\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eCLP,omphalocele, VSD, Single umbilical artery, hyperechoic kidney, NF thickening, EIF\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eTOP\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=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e24\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e47,XY,+18 /Edward's syndrome\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eCLP, VSD, microtia, single umbilical artery\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eTOP\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=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e29\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e47,XY,+12[4]/46,XY[42]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eCLP, thoraco-gastroschisis, cranial coloboma with meningoencephalocele, ectopia cordis, VSD, double outlet right ventricle, pulmonary artery stenosis, strephenopodia, abnormal blood flow spectrum of middle cerebral artery, umbilical artery\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eTOP\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=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e24\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e47,XX,+18 /Edward's syndrome\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eCLP, bilateral choroid plexus cysts, strawberry head, small jaw, microtia, VSD, mild mitral regurgitation, hyperechoic kidney, overlapping fingers\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eTOP\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=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e23\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e47,XY,+13 /Patau's syndrome\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eCLP, agenesis of the corpus callosum or lobate holoprosencephalon, omphalocele, VSD, bilateral kidney enlargement, NF thickening\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eTOP\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=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e34\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e47,XY,+13 /Patau's syndrome\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eCLP, VSD, aortic stenosis, absence of the inferior vermis of the cerebellum, posterior fossa pool broadening, EIF\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eTOP\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=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e18\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e47,XY,+18/Edward's syndrome\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eCLP, VSD, aortic stenosis\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eTOP\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=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e24\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e46,XY,rob(13)(q10;q10) /Patau's syndrome\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eCLP, holoprosencephaly, hypertelorism, flat nose, omphalocele\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eTOP\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=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e24\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e47,XY,+13 /Patau's syndrome\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eVSD, Tetralogy of Fallot, tricuspid regurgitation, hypertelorism, microphthalmia, omphalocele, left polycystic kidney, left ureteral dilatation, left ureterocele\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eTOP\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=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e21\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e46,XY,der(22)(q10;q10)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eCLP, FGR, enlarged cardiothoracic ratio\u003c/p\u003e \u003cp\u003ehypoplasia of the inferior cerebellar vermis\u003c/p\u003e \u003cp\u003eoligohydramnios\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eTOP\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=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e27\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e46,XY,der(13;14),+13 /Patau's syndrome\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eCLP, holoprosencephaly, VSD, aortic stenosis, omphalocele, sacrococcygeal malformations\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eTOP\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=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e28\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e46,XY,der(22)(q10;q10)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eCLP, FGR, abnormal blood flow in the middle cerebral artery, arachnoid cyst, left-sided microphthalmia, micrognathia, cervical lymphatic cystic hygroma,situs inversus\u003c/p\u003e \u003cp\u003ecardiac abnormalities (dextrocardia, pulmonary artery stenosis), low conus medullaris\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eTOP\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=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e31\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e47,XY,+13 /Patau's syndrome\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eCL, holoprosencephaly, EIF\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eTOP\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=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e21\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e47,XY,+13 /Patau's syndrome\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eCLP, omphalocele, polydactyly and overlapping fingers on the right hand, NF thickening \u003c/p\u003e \u003cp\u003eHypoplasia of the cerebellar vermis,Hypoplastic left heart, aortic stenosis, EIF\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eTOP\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=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e29\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e47,XX,+13 /Patau's syndrome\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eCLP, ventriculomegaly, cerebellar hypoplasia, absence of the inferior cerebellar vermis, microphthalmia,\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eTOP\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=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e23\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e47,XY,+13 /Patau's syndrome\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eCL, enlarged right ventricle, EIF\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eTOP\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=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e24\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e46,XX,der(4)t(2;4)(p11;p11)dn\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eCLP, FGR, VSD, hypoplastic left heart, mitral atresia, aortic atresia, cervical and dorsal lymphatic cystic hygroma, small kidneys\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eTOP\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=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e30\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e47,XX,+13 /Patau's syndrome\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eCLP, holoprosencephaly, complex cardiac malformations\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eTOP\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=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e27\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e47,XX,+13 /Patau's syndrome\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eCLP, VSD, holoprosencephaly, hypoplastic left heart, double outlet right ventricle, pulmonary artery stenosis, possible duplex right kidney, single umbilical artery\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eTOP\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=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e30\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e46,XX,del(8)(q21.1q22.2)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eCP, NT thinckening, VSD, nasal bone dysplasia\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eTOP\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=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e38\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e47,XX,+i(12)(p10)[7]/46,XX[104]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eCP, thickened skin on the nasal tip, NF thickening,\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eTOP\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=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e24\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e47,XY,+13 /Patau's syndrome\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eCLP, cerebellar malformation, bilateral microphthalmia, microtia, VSD, right subclavian artery anomaly, polydactyly in both hands, overlapping fingers, polydactyly in both feet, low conus medullaris, dysplastic sacral vertebrae\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eTOP\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e24\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e24\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e47,XXX\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eLP, left choroid plexus cyst, conus medullaris at L4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eLB\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003ctfoot\u003e \u003ctr\u003e\u003ctd colspan=\"5\"\u003eCL, Cleft lip; CLP, Cleft lip and palate; CP, Cleft Palate; NT, nuchal translucency; TOP, termination of pregnancy; VSD, ventricular septal defects; FGR, fetal growth restriction; EIF, echogenic intracardiac focus; TOP: termination of pregnancy; LB, live birth\u003c/td\u003e\u003c/tr\u003e \u003c/tfoot\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cdiv id=\"Sec8\" class=\"Section2\"\u003e \u003ch2\u003eSubmicroscopic aberrations detected by SNP array analysis\u003c/h2\u003e \u003cp\u003eAmong the 138 cases that underwent both karyotyping and SNP array analysis, submicroscopic chromosomal aberrations were identified in 12 cases, including 4 cases with clinically significant submicroscopic abnormalities in individuals with normal karyotypes. The detection rates of additional clinically significant submicroscopic abnormalities were 2.0% (2/101) in the isolated group and 5.4% (2/37) in the non-isolated group (p\u0026thinsp;\u0026gt;\u0026thinsp;0.05). Detailed results are presented in Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\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\u003eSubmicroscopic chromosomal aberrations detected by SNP array analysis.\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"6\"\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 \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCase number\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eUltrasound findings\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eSNP array analysis\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eParental inheritance\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eClinical significance\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003ePregnancy outcome\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e25\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eCL, left echogenic intracardiac focus\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003earr[GRCh37] 22q11.22q11.23(22997929\u0026ndash;25041592)\u0026times;3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eNA\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eVOUS\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eLB\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e26\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eCLP\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003earr[GRCh37] 22q12.3(33,854,040\u0026thinsp;\u0026minus;\u0026thinsp;34,400,183)\u0026times;3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eNA\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eVOUS\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eNA\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e27\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eCP\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003earr[GRCh37] 8q24.22q24.3(134,714,740\u0026thinsp;\u0026minus;\u0026thinsp;146,292,734)\u0026times;2 hmz\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e/\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eVOUS\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eLB\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e28\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eCLP, craniosynostoses\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003earr[GRCh37] Xp22.33(288,066\u0026thinsp;\u0026minus;\u0026thinsp;1,023,657)\u0026times;1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003edn\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003ePathogenic\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eTOP\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e29\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eCP, tricuspid regurgitation\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003earr[GRCh37] 5q23.3q31.1(129,930550-131,441,100)\u0026times;3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eMaternal\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eLikely benign\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eLB\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e30\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eCP\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003earr[GRCh37] 15q11.2q13.1(23693931 28526905)\u0026times;3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003edn\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003ePathogenic\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eTOP\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e31\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eCLP, Ventricular septal defects\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003earr[GRCh37] 4q35.1q35.2(186031030\u0026ndash;189191908)\u0026times;3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003ePaternal\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eVOUS\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eLB\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e32\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eCLP\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003earr[GRCh37] 15q22.33q25.1(67432515\u0026ndash;78796221)\u0026times;2 hmz\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e/\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eVOUS\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eTOP\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e33\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eCLP, VSD, thymusv dysplasia\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003earr[GRCh37] 22q11.21(18916843\u0026ndash;21464764)\u0026times;1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003edn\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003ePathogenic\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eTOP\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e34\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eCLP\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003earr[GRCh37] 6p25.3p25.2(993944_2513338)\u0026times;3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003ePaternal\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eVOUS\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eLB\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e35\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eCP\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003earr[GRCh37] 7q11.23(72669481_74154209)\u0026times;1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eMaternal\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003ePathogenic\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eLB\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e36\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eCLP\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003earr[GRCh37] 5q21.3(106340917\u0026ndash;107886317)\u0026times;1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e/\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eVOUS\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eTOP\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003ctfoot\u003e \u003ctr\u003e\u003ctd colspan=\"6\"\u003eCL, Cleft lip; CLP, Cleft lip and palate; CP, Cleft Palate; NA, not available; VOUS, variants of uncertain significance; TOP, termination of pregnancy; VSD, ventricular septal defects; dn, De novo, LB, live birth\u003c/td\u003e\u003c/tr\u003e \u003c/tfoot\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003c/div\u003e\n\u003ch3\u003ePregnancy outcome\u003c/h3\u003e\n\u003cp\u003ePregnancy outcome data were available for 188 cases (91.7%). Of the 24 pregnancies with clinically significant aberrations, all were terminated except for case 35 (Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e). This fetus presented with cleft palate and a 1.48 Mb deletion at 7q11.23, inherited from a mother with intellectual disability. Among the remaining 160 cases, the termination rate was significantly higher in the non-isolated group at 80.8% (21/26) compared to 23.0% (31/135) in the isolated OFC group (p\u0026thinsp;\u0026lt;\u0026thinsp;0.05). Within the isolated group without clinically significant aberrations, the termination rate for CLP was 39.0% (30/77), which was significantly higher (p\u0026thinsp;\u0026lt;\u0026thinsp;0.05) than that for CL at 11.8% (4/34) and for CP at 3.8% (1/26).\u003c/p\u003e \u003cp\u003eFollow-up data on subsequent pregnancies were obtained for 45 cases, and no recurrent OFC were observed.\u003c/p\u003e"},{"header":"Discussion","content":"\u003cp\u003eThe distribution of different types of OFCs varied across studies \u003csup\u003e[\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e, \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e, \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e]\u003c/sup\u003e. In our study, CLP was more frequently observed than CL or CP. In our study, CLP was more frequently observed than CL or CP. OFCs can occur either in isolation or in combination with other system malformations. Notably, over two-thirds of the cases in this study were isolated OFCs. Since CMA was not widely applied in prenatal diagnosis at our center before 2016, a portion of fetuses with OFCs underwent only conventional karyotyping. In isolated OFCs, no chromosomal abnormalities were detected by conventional karyotyping. By contrast, the chromosomal abnormality rate was as high as 46.2% in the non-isolated group. Consistent with previous studies \u003csup\u003e[\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e, \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e]\u003c/sup\u003e, there was no significant difference in the chromosomal abnormality rates between cases with single-system malformations and those with multisystem malformations. This finding emphasizes that the risk of chromosomal abnormalities is considerably higher when OFCs are accompanied by other structural malformations.\u003c/p\u003e \u003cp\u003eTrisomy 13 has been reported as the most frequent chromosomal abnormality in OFC cases, followed by trisomy 18 \u003csup\u003e[\u003cspan additionalcitationids=\"CR21 CR22\" citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e]\u003c/sup\u003e. Consistent with these findings, our study showed that trisomy 13 accounted for 50.0% of karyotype-detectable abnormalities, followed by trisomy 18 (5 cases, 21.7%) and trisomy 22 (2 cases, 8.7%). This underscores the diagnostic utility of conventional karyotyping in identifying genetic etiologies when OFCs are associated with additional major malformations. Additionally, two cases of low-level mosaicism involving chromosome 12 were detected in this study. Mosaic trisomy 12 is a rare condition, with few prenatal and postnatal cases reported in the literature. Previous studies have highlighted a spectrum of ultrasound findings, from normal to severe congenital anomalies, in pregnancies with trisomy 12 mosaicism \u003csup\u003e[\u003cspan additionalcitationids=\"CR25 CR26\" citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e]\u003c/sup\u003e. One of our cases exhibited 8.9% (4/45) mosaicism in cultured amniocytes and presented with CLP accompanied by multisystem defects involving cardiovascular, abdominal, cranial, and skeletal development. The discordance between low-level mosaicism and severe ultrasound abnormalities might result from the differential distribution of abnormal cells in various tissues, although further testing was unavailable to confirm this hypothesis. Another case involved mosaicism with an extra isochromosome 12p (i(12p)), responsible for Pallister-Killian syndrome (PKS, OMIM: #601803). This case has been detailed in a previous publication \u003csup\u003e[\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e]\u003c/sup\u003e.\u003c/p\u003e \u003cp\u003eIn recent years, chromosomal microarray analysis (CMA) has confirmed the significant role of CNVs in the genetic etiology of OFCs \u003csup\u003e[\u003cspan additionalcitationids=\"CR30 CR31\" citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e]\u003c/sup\u003e. In this study, SNP array analysis identified four pathogenic/likely pathogenic CNVs (2.9%), a detection rate notably lower than postnatal reports \u003csup\u003e[\u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e, \u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e]\u003c/sup\u003e. The 22q11.2 deletion syndrome, associated with DiGeorge syndrome (OMIM: #188400) and velocardiofacial syndrome (OMIM: #192430), is the most common syndrome linked to conotruncal anomalies \u003csup\u003e[\u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e34\u003c/span\u003e, \u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e35\u003c/span\u003e]\u003c/sup\u003e. Mechanistically, haploinsufficiency of \u003cem\u003eTBX1\u003c/em\u003e, encoding a T-box transcription factor essential for pharyngeal arch development, disrupts cranial neural crest cell migration, leading to defective palatal shelf elevation and thymic hypoplasia. A study by Bashir et al. recommended routine 22q11 fluorescence in situ hybridization (FISH) testing for children with CL and/or CP due to the diagnostic implications of velocardiofacial syndrome \u003csup\u003e[\u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e]\u003c/sup\u003e. In present study, we identified a 2.5 Mb microdeletion at 22q11.2 in a fetus with CP, heart defects and thymus gland dysplasia, provides critical prenatal validation of this association. Additionally, a de novo 4.8 Mb at 15q11.2-q13.1 was detected in a fetus with isolated CP. This region encompasses the critical region of 15q11-q13 duplication syndrome (OMIM: #608636), which is typically characterized by neurodevelopmental disorders (autism, epilepsy) and hypotonia\u003csup\u003e[\u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e37\u003c/span\u003e]\u003c/sup\u003e. To the best of our knowledge, this duplication has not been previously reported in cases of OFCs, which expands the phenotypic spectrum of 15q11-q13 CNVs and highlights potential genotype-phenotype correlations requiring further validation. In a male fetus with CLPand craniosynostoses, a 736 Kb deletion was identified at Xp22.33, involving the Short Stature Homeobox (SHOX) gene. Haploinsufficiency of SHOX is primarily associated with L\u0026eacute;ri-Weill dyschondrosteosis\u003csup\u003e[\u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e38\u003c/span\u003e, \u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e39\u003c/span\u003e]\u003c/sup\u003e and facial dysmorphism has been observed in some cases with Xp22.33 microdeletions\u003csup\u003e[\u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e40\u003c/span\u003e, \u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e41\u003c/span\u003e]\u003c/sup\u003e. A study by Chen et al. concluded that SHOX2 plays a critical role in osteogenesis of a specific cell lineage and contributes to the development of the palatine process of the maxilla by interacting with distal cis-regulatory elements to regulate skeletogenic gene expression and pattern the hard palate \u003csup\u003e[\u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e42\u003c/span\u003e]\u003c/sup\u003e. Therefore, the relationship between OFCs and Shox is worthy of further in-depth study. Furthermore, a 1.5 Mb deletion at 7q11.23, responsible for Williams-Beuren syndrome (WBS; OMIM: #194050), was identified in a fetus with isolated CP. The most common manifestations of WBS include a characteristic craniofacial morphology, developmental delays, cognitive and behavioural changes\u003csup\u003e[\u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e43\u003c/span\u003e]\u003c/sup\u003e. Reports of WBS patients with cleft palate are not uncommon\u003csup\u003e[\u003cspan additionalcitationids=\"CR45 CR46\" citationid=\"CR44\" class=\"CitationRef\"\u003e44\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR47\" class=\"CitationRef\"\u003e47\u003c/span\u003e]\u003c/sup\u003e, suggesting partial phenotypic overlap with this congenital anomaly. Genetic analysis confirmed maternal inheritance of the deletion. While the mother\u0026rsquo;s primary manifestation was intellectual disability, the fetus developed CP and may face heightened risks of multisystem involvement due to allelic dosage effects or variable expressivity of the 7q11.23 deletion. Following comprehensive genetic counseling regarding prognosis and recurrence risks, the family opted to continue the pregnancy.\u003c/p\u003e \u003cp\u003eRegarding pregnancy outcomes, genetic abnormalities of clinical significance and non-isolated OFCs were the primary reasons for pregnancy termination. Additionally, we placed special attention to isolated OFCs. CLP cases had the highest termination rate, whereas all 22 cases of isolated CP were continued. Notably, follow-up of 45 cases revealed no recurrence of OFCs in subsequent pregnancies. These findings suggest that karyotyping and SNP array analysis are effective in identifying the genetic etiologies of primary OFCs, thereby providing valuable guidance for future pregnancies.\u003c/p\u003e \u003cp\u003eThis study has several limitations. Its retrospective design and the fact that not all cases underwent SNP array analysis may have introduced bias in evaluating the incidence of CNVs and single-gene disorders in fetuses with OFCs.\u003c/p\u003e \u003cp\u003eIn conclusion, this study underscores the effectiveness of comprehensive genetic evaluations, including karyotyping and SNP array analysis, in pregnancies with first-occurrence OFCs. Notably, chromosomal karyotype abnormalities are unlikely to cause isolated OFCs. Moreover, pregnancy outcomes for OFCs are primarily determined by the chromosomal analysis results, the specific type of OFCs, and the presence of other systemic malformation.\u003c/p\u003e"},{"header":"Abbreviations","content":"\u003cdiv class=\"DefinitionList\"\u003e \u003cdiv class=\"DefinitionListEntry\"\u003e \u003cdiv class=\"Term\"\u003eOFCs\u003c/div\u003e \u003cdiv class=\"Description\"\u003e \u003cp\u003eOrofacial clefts\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv class=\"DefinitionListEntry\"\u003e \u003cdiv class=\"Term\"\u003eCL\u003c/div\u003e \u003cdiv class=\"Description\"\u003e \u003cp\u003ecleft lip\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv class=\"DefinitionListEntry\"\u003e \u003cdiv class=\"Term\"\u003eCP\u003c/div\u003e \u003cdiv class=\"Description\"\u003e \u003cp\u003ecleft palate\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv class=\"DefinitionListEntry\"\u003e \u003cdiv class=\"Term\"\u003eCLP\u003c/div\u003e \u003cdiv class=\"Description\"\u003e \u003cp\u003ecleft lip and palate\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv class=\"DefinitionListEntry\"\u003e \u003cdiv class=\"Term\"\u003eCNVs\u003c/div\u003e \u003cdiv class=\"Description\"\u003e \u003cp\u003ecopy number variations\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv class=\"DefinitionListEntry\"\u003e \u003cdiv class=\"Term\"\u003eISCN\u003c/div\u003e \u003cdiv class=\"Description\"\u003e \u003cp\u003eInternational System for Human Cytogenetic Nomenclature\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv class=\"DefinitionListEntry\"\u003e \u003cdiv class=\"Term\"\u003eCHAS\u003c/div\u003e \u003cdiv class=\"Description\"\u003e \u003cp\u003eChromosome Analysis Suite\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv class=\"DefinitionListEntry\"\u003e \u003cdiv class=\"Term\"\u003eACMG\u003c/div\u003e \u003cdiv class=\"Description\"\u003e \u003cp\u003eAmerican College of Medical Genetics\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv class=\"DefinitionListEntry\"\u003e \u003cdiv class=\"Term\"\u003eCMA\u003c/div\u003e \u003cdiv class=\"Description\"\u003e \u003cp\u003echromosomal microarray analysis\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv class=\"DefinitionListEntry\"\u003e \u003cdiv class=\"Term\"\u003eFISH\u003c/div\u003e \u003cdiv class=\"Description\"\u003e \u003cp\u003efluorescence in situ hybridization\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv class=\"DefinitionListEntry\"\u003e \u003cdiv class=\"Term\"\u003eSHOX\u003c/div\u003e \u003cdiv class=\"Description\"\u003e \u003cp\u003eShort Stature Homeobox\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv class=\"DefinitionListEntry\"\u003e \u003cdiv class=\"Term\"\u003eWBS\u003c/div\u003e \u003cdiv class=\"Description\"\u003e \u003cp\u003eWilliams-Beuren syndrome\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003c/div\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eEthics approval and consent to participate\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe present study was approved by the Protection of Human Ethics Committee of Fujian Maternity and Child Health Hospital. Written informed consent was obtained from each pregnant woman.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent for publication\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eWritten informed consent to publish this case was obtained from all the participants and parents of the minor participants, including case description and medical data.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAvailability of data and material\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCompeting interests\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors declare they have no conflict of interest.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe study was supported by Key Special Projects of Fujian Provincial Department of Science and Technology (grant No. 2021YZ034011). Innovation Platform Project of Science and Technology, Fujian province (2021Y2012) National Key Clinical Specialty Construction Program of China (Obstetric).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthors' contributions\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eXW and XX drafted the main manuscript. JL, LS and BL prepared Tables 1–3,\u0026nbsp;NL\u0026nbsp;and LX provided genetic counseling.\u0026nbsp;DG, LZreviewed and revised the manuscript. All authors reviewed the manuscript.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAcknowledgements\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eVyas T, P Gupta, S Kumar, R Gupta, T Gupta, and H P Singh, Cleft of lip and palate: A review. J Family Med Prim Care, 2020; 9(6): p. 2621-2625.\u003c/li\u003e\n\u003cli\u003eSalari N, N Darvishi, M Heydari, S Bokaee, F Darvishi, and M Mohammadi, Global prevalence of cleft palate, cleft lip and cleft palate and lip: A comprehensive systematic review and meta-analysis. J Stomatol Oral Maxillofac Surg, 2022; 123(2): p. 110-120.\u003c/li\u003e\n\u003cli\u003eWang M, Y Yuan, Z Wang, D Liu, Z Wang, F Sun, et al., Prevalence of Orofacial Clefts among Live Births in China: A Systematic Review and Meta-Analysis. Birth Defects Res, 2017; 109(13): p. 1011-1019.\u003c/li\u003e\n\u003cli\u003eTonni G, G Grisolia, and W Sepulveda, Early prenatal diagnosis of orofacial clefts: evaluation of the retronasal triangle using a new three-dimensional reslicing technique. Fetal Diagn Ther, 2013; 34(1): p. 31-7.\u003c/li\u003e\n\u003cli\u003ePekar-Zlotin M, N Zilberman Sharon, Y Melcer, Y Tal-Bliman, J Ezratty, M Feingold-Zadok, et al., Pregnancy with Facial Cleft: 20 Years of Experience at a Single Center. Isr Med Assoc J, 2023; 25(10): p. 678-682.\u003c/li\u003e\n\u003cli\u003eExpert Group of Multidisciplinary Treatment Process of Cleft L and Palate, [Specification of the multidisciplinary treatment process of cleft lip and palate during pregnancy, prenatal and postnatal stages]. Zhonghua Kou Qiang Yi Xue Za Zhi, 2021; 56(11): p. 1059-1065.\u003c/li\u003e\n\u003cli\u003eBeaty T H, M L Marazita, and E J Leslie, Genetic factors influencing risk to orofacial clefts: today\u0026apos;s challenges and tomorrow\u0026apos;s opportunities. F1000Res, 2016; 5: p. 2800.\u003c/li\u003e\n\u003cli\u003eBabai A and M Irving, Orofacial Clefts: Genetics of Cleft Lip and Palate. Genes (Basel), 2023; 14(8).\u003c/li\u003e\n\u003cli\u003eSchindewolf E, N Khalek, M P Johnson, J Gebb, B Coleman, T B Crowley, et al., Expanding the fetal phenotype: Prenatal sonographic findings and perinatal outcomes in a cohort of patients with a confirmed 22q11.2 deletion syndrome. Am J Med Genet A, 2018; 176(8): p. 1735-1741.\u003c/li\u003e\n\u003cli\u003eLi S, A Chao, Z Li, C A Moore, Y Liu, J Zhu, et al., Folic acid use and nonsyndromic orofacial clefts in China: a prospective cohort study. Epidemiology, 2012; 23(3): p. 423-32.\u003c/li\u003e\n\u003cli\u003eSalazar Trujillo A, C Rinc\u0026oacute;n-Guio, L L\u0026oacute;pez Narv\u0026aacute;ez, J C\u0026aacute;ceres, and J D Charry, First trimester sonographic diagnosis of orofacial defects. Review of literature. J Matern Fetal Neonatal Med, 2020; 33(18): p. 3200-3206.\u003c/li\u003e\n\u003cli\u003eFan K L, C K Black, E Mantilla-Rivas, D I Bulas, E Rubio, A R Blask, et al., Coordination of the Fetal Medicine Institute and the Cleft and Craniofacial Center: Application to Early Management of Infants With Cleft Lip and Palate. J Craniofac Surg, 2019; 30(7): p. 2061-2064.\u003c/li\u003e\n\u003cli\u003eWu X, Y Li, L Su, X Xie, M Cai, N Lin, et al., Chromosomal Microarray Analysis for the Fetuses with Aortic Arch Abnormalities and Normal Karyotype. Mol Diagn Ther, 2020; 24(5): p. 611-619.\u003c/li\u003e\n\u003cli\u003eWu X, L Xu, Y Li, N Lin, L Su, M Cai, et al., Submicroscopic aberrations of chromosome 16 in prenatal diagnosis. Mol Cytogenet, 2019; 12: p. 36.\u003c/li\u003e\n\u003cli\u003eMcgowan-Jordan J, R J Hastings, and S Moore, An International System for Human Cytogenomic Nomenclature (2020). Cytogenetic and genome research, 2020.\u003c/li\u003e\n\u003cli\u003eSouth S T, C Lee, A N Lamb, A W Higgins, H M Kearney, G Working Group for the American College of Medical, et al., ACMG Standards and Guidelines for constitutional cytogenomic microarray analysis, including postnatal and prenatal applications: revision 2013. Genet Med, 2013; 15(11): p. 901-9.\u003c/li\u003e\n\u003cli\u003eLee C W, S M Hwang, Y S Lee, M A Kim, and K Seo, Prevalence of orofacial clefts in Korean live births. Obstet Gynecol Sci, 2015; 58(3): p. 196-202.\u003c/li\u003e\n\u003cli\u003eBeriaghi S, S Myers, S Jensen, S Kaimal, C Chan, and G B Schaefer, Cleft lip and palate: association with other congenital malformations. Journal of Clinical Pediatric Dentistry, 2009; 33(3): p. 207-210.\u003c/li\u003e\n\u003cli\u003eAzadgoli B, N C O Munabi, A Fahradyan, A Auslander, M McCullough, N Aflatooni, et al., Congenital Heart Disease in Patients With Cleft Lip/Palate and Its Impact on Cleft Management. Cleft Palate Craniofac J, 2020; 57(8): p. 957-966.\u003c/li\u003e\n\u003cli\u003ePetry P, J B Polli, V F Mattos, R C Rosa, P R Zen, C Graziadio, et al., Clinical features and prognosis of a sample of patients with trisomy 13 (Patau syndrome) from Brazil. Am J Med Genet A, 2013; 161a(6): p. 1278-83.\u003c/li\u003e\n\u003cli\u003eBerg\u0026eacute; S J, H Plath, P T Van de Vondel, T Appel, B Niederhagen, J J Von Lindern, et al., Fetal cleft lip and palate: sonographic diagnosis, chromosomal abnormalities, associated anomalies and postnatal outcome in 70 fetuses. Ultrasound Obstet Gynecol, 2001; 18(5): p. 422-31.\u003c/li\u003e\n\u003cli\u003eBruns D, Birth history, physical characteristics, and medical conditions in long-term survivors with full trisomy 13. Am J Med Genet A, 2011; 155a(11): p. 2634-40.\u003c/li\u003e\n\u003cli\u003eVibert F, G Schmidt, K L\u0026ouml;ffler, A Gasiorek-Wiens, W Henrich, and S Verlohren, Accuracy of prenatal detection of facial clefts and relation between facial clefts, additional malformations and chromosomal abnormalities: a large referral-center cohort. Arch Gynecol Obstet, 2024; 309(5): p. 1971-1980.\u003c/li\u003e\n\u003cli\u003eChen C P, Y N Su, J W Su, S R Chern, Y T Chen, L F Chen, et al., Mosaic trisomy 12 at amniocentesis: prenatal diagnosis and molecular genetic analysis. Taiwan J Obstet Gynecol, 2013; 52(1): p. 97-105.\u003c/li\u003e\n\u003cli\u003eChen C P, C J Lin, S R Chern, P S Wu, Y N Chen, S W Chen, et al., Prenatal diagnosis and molecular cytogenetic characterization of low-level mosaic trisomy 12 at amniocentesis associated with a favorable pregnancy outcome. Taiwan J Obstet Gynecol, 2017; 56(2): p. 238-242.\u003c/li\u003e\n\u003cli\u003eRoberts W, A Zurada, A Zurada-ZieliŃSka, J Gielecki, and M Loukas, Anatomy of trisomy 12. Clin Anat, 2016; 29(5): p. 633-7.\u003c/li\u003e\n\u003cli\u003eHong B, J Zunich, A Openshaw, and R M Toydemir, Clinical features of trisomy 12 mosaicism-Report and review. Am J Med Genet A, 2017; 173(6): p. 1681-1686.\u003c/li\u003e\n\u003cli\u003eWu X, X Xie, L Su, N Lin, B Liang, N Guo, et al., Prenatal diagnosis of Pallister-Killian syndrome and literature review. J Cell Mol Med, 2021; 25(18): p. 8929-8935.\u003c/li\u003e\n\u003cli\u003eLansdon L A, B W Darbro, A L Petrin, A M Hulstrand, J M Standley, R B Brouillette, et al., Identification of Isthmin 1 as a Novel Clefting and Craniofacial Patterning Gene in Humans. Genetics, 2018; 208(1): p. 283-296.\u003c/li\u003e\n\u003cli\u003eCai Y, K E Patterson, F Reinier, S E Keesecker, E Blue, M Bamshad, et al., Copy Number Changes Identified Using Whole Exome Sequencing in Nonsyndromic Cleft Lip and Palate in a Honduran Population. Birth Defects Res, 2017; 109(16): p. 1257-1267.\u003c/li\u003e\n\u003cli\u003eConte F, M Oti, J Dixon, C E Carels, M Rubini, and H Zhou, Systematic analysis of copy number variants of a large cohort of orofacial cleft patients identifies candidate genes for orofacial clefts. Hum Genet, 2016; 135(1): p. 41-59.\u003c/li\u003e\n\u003cli\u003eLei T Y, H T Wang, F Li, Y Q Cui, F Fu, R Li, et al., Application of high resolution SNP arrays in patients with congenital oral clefts in south China. J Genet, 2016; 95(4): p. 801-809.\u003c/li\u003e\n\u003cli\u003eda Silva H P V, G H M Oliveira, M A G Ururahy, J F Bezerra, K S C de Souza, R H Bortolin, et al., Application of high-resolution array platform for genome-wide copy number variation analysis in patients with nonsyndromic cleft lip and palate. J Clin Lab Anal, 2018; 32(6): p. e22428.\u003c/li\u003e\n\u003cli\u003eMcDonald-McGinn D M and K E Sullivan, Chromosome 22q11.2 deletion syndrome (DiGeorge syndrome/velocardiofacial syndrome). Medicine (Baltimore), 2011; 90(1): p. 1-18.\u003c/li\u003e\n\u003cli\u003eLewyllie A, J Roosenboom, K Indencleef, P Claes, A Swillen, K Devriendt, et al., A Comprehensive Craniofacial Study of 22q11.2 Deletion Syndrome. J Dent Res, 2017; 96(12): p. 1386-1391.\u003c/li\u003e\n\u003cli\u003eBashir M A, P D Hodgkinson, T Montgomery, and M Splitt, 22q11 Deletion in children with cleft lip and palate--is routine screening justified? J Plast Reconstr Aesthet Surg, 2008; 61(2): p. 130-2.\u003c/li\u003e\n\u003cli\u003eUrraca N, J Cleary, V Brewer, E K Pivnick, K McVicar, R L Thibert, et al., The interstitial duplication 15q11.2-q13 syndrome includes autism, mild facial anomalies and a characteristic EEG signature. Autism Res, 2013; 6(4): p. 268-79.\u003c/li\u003e\n\u003cli\u003eShima H, T Tanaka, T Kamimaki, S Dateki, K Muroya, R Horikawa, et al., Systematic molecular analyses of SHOX in Japanese patients with idiopathic short stature and Leri-Weill dyschondrosteosis. J Hum Genet, 2016; 61(7): p. 585-91.\u003c/li\u003e\n\u003cli\u003eVannelli S, M Baffico, R Buganza, F Verna, G Vinci, D Tessaris, et al., SHOX deficiency in children with growth impairment: evaluation of known and new auxological and radiological indicators. Ital J Pediatr, 2020; 46(1): p. 163.\u003c/li\u003e\n\u003cli\u003eDepeyre A, M Schlund, R Nicot, and J Ferri, Dental and Maxillofacial Signs in Leri-Weill Dyschondrosteosis. J Oral Maxillofac Surg, 2019; 77(4): p. 762-768.\u003c/li\u003e\n\u003cli\u003eDupont C, A Lebbar, C Teinturier, F Baverel, G Viot, D Le Tessier, et al., First reported case of intrachromosomal cryptic inv dup del Xp in a boy with developmental retardation. Am J Med Genet A, 2007; 143a(11): p. 1236-43.\u003c/li\u003e\n\u003cli\u003eXu J, L Wang, H Li, T Yang, Y Zhang, T Hu, et al., Shox2 regulates osteogenic differentiation and pattern formation during hard palate development in mice. J Biol Chem, 2019; 294(48): p. 18294-18305.\u003c/li\u003e\n\u003cli\u003eKozel B A, B Barak, C A Kim, C B Mervis, L R Osborne, M Porter, et al., Williams syndrome. Nat Rev Dis Primers, 2021; 7(1): p. 42.\u003c/li\u003e\n\u003cli\u003eBlanco-D\u0026aacute;vila F and J A Olveda-Rodriguez, Cleft palate in a patient with Williams\u0026apos; syndrome. J Craniofac Surg, 2001; 12(2): p. 145-7.\u003c/li\u003e\n\u003cli\u003eDomenico S, C Orlando, F F Graziana, P Papi, and A Giulia, Cleft palate in Williams syndrome. Ann Maxillofac Surg, 2013; 3(1): p. 84-6.\u003c/li\u003e\n\u003cli\u003eVincent C, J M Mercier, and A David, [Cleft palate and Williams syndrome]. Rev Stomatol Chir Maxillofac, 2008; 109(1): p. 44-7.\u003c/li\u003e\n\u003cli\u003eYamaguchi T, T Shirota, M Adel, M Takahashi, S Haga, R Nagahama, et al., Orthodontic Treatment and Maxillary Anterior Segmental Distraction Osteogenesis of a Subject with Williams-Beuren Syndrome and Isolated Cleft Palate: A Long-Term Follow-Up from the Age of 5 to 24 Years. Case Rep Dent, 2017; 2017: p. 7019045.\u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"orofacial clefts, cleft lip, cleft palate, chromosomal abnormality, isolated, non-isolated","lastPublishedDoi":"10.21203/rs.3.rs-6191104/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-6191104/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003ch2\u003eBackground\u003c/h2\u003e \u003cp\u003eOrofacial clefts (OFCs) are among the most common birth defects. This study aimed to investigate the prenatal genetic evaluation and pregnancy outcomes of pregnancies with first-occurrence typical OFCs.\u003c/p\u003e\u003ch2\u003eMethods\u003c/h2\u003e \u003cp\u003e We retrospectively reviewed 205 first-occurrence OFCs pregnancies during 2010 and 2024, including cleft lip (CL, n\u0026thinsp;=\u0026thinsp;42), cleft palate (CP, n\u0026thinsp;=\u0026thinsp;31), and cleft lip and palate (CLP, n\u0026thinsp;=\u0026thinsp;132). Based on the presence of additional malformations, cases were categorized as isolated (n\u0026thinsp;=\u0026thinsp;153) or non-isolated (n\u0026thinsp;=\u0026thinsp;52). Conventional karyotyping was used to detect chromosomal abnormalities, and single nucleotide polymorphism array (SNP array) analysis was performed in 138 cases to identify submicroscopic aberrations.\u003c/p\u003e\u003ch2\u003eResults\u003c/h2\u003e \u003cp\u003eThe proportions of isolated cases for CL, CP, and CLP were 95.2% (40/42), 83.8% (26/31), and 65.9% (87/132), respectively. Conventional karyotyping identified chromosomal abnormalities in 24 cases (11.7%), with trisomy 13 being the most common (12 cases, 50.0%), followed by trisomy 18 (5 cases, 20.8%). All abnormalities were observed in the non-isolated group, where the chromosomal abnormality rate was 46.2% (24/52). In this group, the chromosomal abnormality rates for CL, CP, and CLP were 50.0% (1/2), 20.0% (1/5), and 48.9% (22/45), respectively. SNP array analysis in 138 cases revealed clinically significant submicroscopic aberrations in 4 karyotypically normal cases, with incremental detection rates of 2.0% (2/101) in the isolated group and 5.4% (2/37) in the non-isolated group (p\u0026thinsp;\u0026gt;\u0026thinsp;0.05). The pregnancy termination rates were 25.5% (35/137) for the isolated group and 90.2% (46/51) for the non-isolated group. Among pregnancies with isolated OFCs and no clinically significant submicroscopic or microscopic chromosomal abnormalities, the termination rates were 11.8% (4/34) for CL, 3.8% (1/26) for CP, and 39.0% (30/77), respectively. Longitudinal follow-up of 46 cases with subsequent pregnancies revealed no recurrence of OFC.\u003c/p\u003e\u003ch2\u003eConclusion\u003c/h2\u003e \u003cp\u003eMost fetal OFCs are isolated, presenting a very low risk of chromosomal abnormalities. For pregnancies with first-occurrence OFCs, the integration of karyotyping and SNP array analysis effectively evaluates the genetic etiology and provides essential guidance for prenatal counseling and future pregnancy management.\u003c/p\u003e","manuscriptTitle":"Prenatal genetic evaluation and outcomes in pregnancies with first-occurrence typical orofacial clefts","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-03-31 06:43:35","doi":"10.21203/rs.3.rs-6191104/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"14fea98c-f1c9-41ce-a53c-039224d7172b","owner":[],"postedDate":"March 31st, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2025-04-15T11:38:24+00:00","versionOfRecord":[],"versionCreatedAt":"2025-03-31 06:43:35","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-6191104","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-6191104","identity":"rs-6191104","version":["v1"]},"buildId":"XKTyCvWXoU3ODBz1xrDgd","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}