Molecular Etiology and Epidemiological Features of a Sapovirus GI.6 Diarrheal Disease Cluster in Jiangxi, China

Molecular Etiology and Epidemiological Features of a Sapovirus GI.6 Diarrheal Disease Cluster in Jiangxi, China | 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 Article Molecular Etiology and Epidemiological Features of a Sapovirus GI.6 Diarrheal Disease Cluster in Jiangxi, China Qian Wang, Jun Zhou, Shiwen Liu, Yanni Zhang, Hailong Liu, Xiaoqing Liu, and 9 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7881481/v1 This work is licensed under a CC BY 4.0 License Status: Under Review Version 1 posted 12 You are reading this latest preprint version Abstract Objective To clarify the etiological and genetic evolutionary characteristics of an acute gastroenteritis cluster epidemic caused by sapovirus in Nanchang, Jiangxi, China. Methods A total of 10 anal swab samples were collected from student and teacher cases during a gastroenteritis epidemic at a primary school in Nanchang, Jiangxi Province in November 2024. Real-time fluorescent RT-PCR was used for routine gene detection of diarrhea viruses. For the sapovirus nucleic acid-positive samples, traditional RT-PCR was used for amplification, sequence determination, splicing, and genetic evolutionary analysis. Results This epidemic cluster was formed due to contact transmission. Real-time fluorescent RT-PCR detection confirmed that 5 samples were positive for sapovirus nucleic acid, and the capsid protein gene VP1 fragments were successfully amplified. Genetic evolutionary analysis revealed that the molecular type of sapovirus was the GI.6 genotype, with 100% homology among the 5 sequences. The nucleotide homology between the sequences in this study and the sapovirus GI.6 reference sequence ranged from 97.07% to 99.12%, and the amino acid homology ranged from 96.49% to 97.37%. Amino acid variation site analysis revealed that there were 4 amino acid site substitutions between them. The closest distance of nucleotide homology was 99.12% from Zhejiang, China in 2023 (OR351089)。 Conclusion The causative agent of this epidemic cluster was identified as sapovirus GI.6, a genotype rarely reported in Jiangxi. In the future, further research on the molecular epidemiology of sapovirus with more gene sites that may mutate should be strengthened. Health sciences/Diseases Biological sciences/Genetics Biological sciences/Microbiology Biological sciences/Molecular biology Sapovirus acute gastroenteritis epidemic cluster molecular epidemiology Figures Figure 1 Figure 2 Introduction Sapovirus (SaV) belongs to the genus Sapovirus in the family Caliciviridae and is one of the important pathogens that cause human acute gastroenteritis (AGE) [ 1 ] . SaV is a single-stranded positive-sense RNA virus with a genome of approximately 7.1–7.6 kb, and is divided into 2–3 open reading frames (ORFs). ORF1 encodes six non-structural proteins NSP1 ~ NSP7 and one major capsid structural protein VP1. ORF2 encodes the smaller structural protein VP2. The function of ORF3 remains unclear [ 2 ] . According to the nucleotide sequence differences of the capsid protein VP1 encoded by ORF1, SaV can be divided into 20 genotypes (GI-GXX), among which GI, GII, GIV, and GV can infect humans [ 3 ] . Human SaV is mainly transmitted through the fecal-oral route. Its clinical symptoms include dysphoria, nausea, vomiting, diarrhea, abdominal pain, and abdominal colic. According to sentinel surveillance data at both China national and Jiangxi provincial levels, sporadic cases occur throughout the year, with peak incidence observed during winter and spring seasons. It is characterized by diverse transmission routes, a low infectious dose, and general susceptibility of the population. Gastroenteritis induced by SaV is usually milder than that caused by NoV and is relatively rare in Jiangxi Province. However, with the widespread use of the rotavirus vaccine, outbreaks of acute gastroenteritis (AGE) caused by SaV in kindergartens, nursing homes, and schools are increasingly becoming a major public health issue [ 4 ] . In November 2024, a clustered outbreak of acute gastroenteritis occurred in a primary school in Nanchang, Jiangxi. This study conducted pathogen identification for this outbreak, as well as sequencing typing and genetic evolution analysis, aiming to understand the evolutionary characteristics of SaV and provide more effective strategies and means for the prevention and control of viral diarrhea. Methods Sample collection According to the definitions of suspected cases, clinically diagnosed cases, and confirmed cases in the "National Surveillance Program for Viral Diarrhea (2021 Edition)", a suspected case is defined as any teacher, student, or staff member at the Yuquandao Campus of Nanchang Normal University Affiliated Primary School who has presented with gastrointestinal symptoms such as nausea, vomiting, abdominal pain, and diarrhea since November 5, 2024. A clinically diagnosed case refers to a suspected case that meets the criteria of having ≥3 bowel movements with a change in stool consistency (watery stools) within 24 hours and/or ≥2 episodes of vomiting within 24 hours, without other etiological diagnoses, and having an epidemiological association with a confirmed case. A confirmed case is a suspected or clinically diagnosed case with a positive nucleic acid test result for Sapovirus in fecal, anal swab, or vomitus samples. The Centers for Disease Control and Prevention (CDC) in Qingshanhu District conducted a case search by reviewing the school's morning check records and records of absences due to illness, and interviewing school administrators and class teachers. On November 12, 2024, a total of 10 anal swab samples were collected from suspected cases on-site and sent to the district CDC laboratory for testing. Sample processing and viral nucleic acid extraction After vortexing the collected anal swab samples at 2500 r/min for 2 minutes, let them stand for 5 minutes for later use. A fully automatic nucleic acid extractor (Strean SP96, Guangzhou Daan Gene) and its matching nucleic acid extraction or purification kit (catalog no. DA0602) were used to extract viral nucleic acid according to the kit instructions. The extracted nucleic acid was stored at 4°C for short-term storage and at -80°C for long-term storage. Real-time fluorescent RT-PCR (real-time RT-PCR) Using the extracted viral nucleic acid as a template, perform real-time fluorescent RT-PCR detection of the virus with triple Real-time Fluorescent PCR Detection Kit for Group A Rotavirus/Adenovirus/Astrovirus Nucleic Acid (Beijing Applied Biological Technologies Co., Ltd. Catalog No.: A7133YH-50T) and Triple Real-time Fluorescent PCR Detection Kit for Norovirus Genogroup I/Norovirus Genogroup II/Sapovirus Nucleic Acid (Beijing Applied Biological Technologies Co., Ltd. Catalog No.: A7123YH-50T). The total reaction volume is 25 μL. The amplification conditions and result judgment shall be carried out according to the kit instructions. Amplification and sequencing of the SaV capsid region (VP1) For the samples positive for SaV in 1.3, the primer pair SLV5317 and SLV5749 was used to amplify the SaV VP1 gene [11]. SLV5317: 5’-CTC GCC ACC TAC RAW GCB TGG TT-3’, SLV5749: 5’-CGG RCY TCA AAV STA CCB CCC CA-3’. The reactions were performed using the One-Step RT-PCR Kit (QIAGEN, Germany; Cat. No. 210212) on an Eppendorf thermal cycler. The reaction system was as follows: 8 μL of 5xRT-PCR Buffer, 1.6 μL of dNTP mixture (10 mM) , 1.6 μL of RT-PCR Enzyme mixture,0.8 μL of SLV5317 (30 μM),0.8 μL of SLV5749 (30 μM),12 μL of DEPC-treated H₂O,and 8 μL of RNA template, for a total volume of 40 μL. The PCR conditions were as follows: reverse transcription at 50°C for 30 min; initial denaturation at 95°C for 15 min; 40 cycles of denaturation at 94°C for 30 s, annealing at 50°C for 30 s and extension at 72°C for 1 min, and a final extension at 72°C for 10 min; 4°C indefinitely. The expected PCR product size was 434bp. The PCR products were sent to Sangon Biotechnology (Shanghai) Co., Ltd. for sequencing. Sequence splicing and molecular genetic evolution analysis of SaV VP1 The SeqMan software was used to organize and splice the sequences. At the same time, the sequences of the VP1 region of SaV GI~GVII types were downloaded from GenBank. Using the ClustalW method in the MEGA 12.0 software, the sequences that had been sequenced and spliced were compared with the nucleotide sequences of the VP1 region of SaV downloaded from the GenBank database.The neighbor-joining algorithm and maximum composite likelihood model with 1000 bootstraps method was used to construct a phylogenetic tree. NCBI BLAST was used to analyze the nucleotide and amino acid sequence homology, and Megalin software was used to analyze the average genetic distance of the sequences and the amino acid variation sites. Results Basic situation of the epidemic The primary school where the epidemic occurred has a total of 6 grades, including 64 classes, with a total of 2,890 students and 201 teaching and administrative staff. As of 15:00 on November 20, 2024, a total of 10 suspected cases were found, all of whom were teachers and students in Class 10, Grade 5. The male-to-female ratio of the cases was 2.33:1. Nine cases were students, and 1 case was a teacher. Combining the results of epidemiological investigations and laboratory tests, among them, 5 cases were confirmed cases, and 5 cases were clinically diagnosed cases. The main clinical manifestations of these 10 cases were vomiting, nausea, abdominal pain, and diarrhea, and 2 cases had dysphoria. The earliest onset time of the cases was 11:00 on November 8, and the latest onset time was 9:00 on November 12. The peak onset time was concentrated from 00:00 on November 11 to 12:00 on November 12. Table 1: Statistical table of clinical symptoms Symptoms Number of cases (N=10) Percentage (%) Vomiting 8 80.00 Abdominal pain 8 80.00 Nausea 6 60.00 Diarrhea 3 30.00 Dysphoria 2 20.00 Real-time RT-PCR detection results Among the 10 samples (all were anal swabs from 10 suspected cases), 5 cases were detected positive for SaV, with a positive rate of 50% (5/10), and the others were all negative. Sequencing and analysis of the SaV VP1 All five positive samples detected in Section 3.2 exhibited successful amplification via RT-PCR, with the resulting amplicons matching the predicted fragment length. Forty-five reference sequences of the VP1 region of each typical and representative SaV genotype globally were selected in this study. All the reference strain sequences of each SaV genotype were from the National Center for Biotechnology Information (NCBI). Genetic evolutionary analysis was performed with the 5 sequences in this epidemic cluster, and a phylogenetic tree was constructed. The evolutionary analysis showed that the SaV in this outbreak belonged to SaV GI.6, which was most closely related to the isolate from Zhejiang, China in 2023 (OR351089). The phylogenetic tree is shown in Figure 1. Results of gene nucleotide and amino acid homology analysis Since the sequencing results of the VP1 region of 5 SaV samples in this epidemic showed 100% homology after comparison, one of them (2024FXYQ153) was selected for nucleotide homology analysis with GI.6 in the reference sequence. The results showed that the nucleotide homology between 2024FXYQ153 and the VP1 partial region of the SaV GI.6 reference sequence ranged from 97.07% to 99.12%, and the amino acid homology ranged from 96.49% to 97.37%. See Table 2 for details. Table 2: Results of nucleotide and amino acid homology analysis of the SaV sequence and the reference sequence of the VP1 gene in this study Sample ID/Genbank ID Nucleotide homology （ % ） Amino acid homology （ % ） 2024FXYQ153 100.00 100.00 OR351089/ZJ/CN/2023 99.12 97.37 AB455803/JPN/2005 98.83 96.49 MN509083/US/2016 98.83 96.49 AB6224354/JPN/2008 98.83 96.49 LC380411/JPN/2005 98.83 96.49 MG012443/US/2015 98.53 96.49 OQ835589/SY/CN/2021 98.53 97.37 AJ606694/UK/2000 97.07 97.37 Results of average genetic distance analysis The average nucleotide genetic distance between 2024FXYQ153 and GI.6 in the reference sequences was analyzed. The results showed that the average genetic distance between 2024FXYQ153 and the partial VP1 region of the SaV GI.6 reference sequences ranged from 0 to 0.0330. See Table 3 for details. Table 3 Analysis of the average nucleotide genetic distance of the VP1 gene between the SaV sequences in this study and the reference sequences Sample ID/Genbank ID Sample ID/Genbank ID 2024FXYQ153 OQ835589 OR351089 MG012443 LC380411 AJ606694 AB622435 MN509083 AB455803 2024FXYQ153 OQ835589 0.0178 OR351089 0.0118 0.0177 MG012443 0.0178 0.0118 0.0117 LC380411 0.0148 0.0088 0.0088 0.0029 AJ606694 0.0330 0.0329 0.0298 0.0267 0.0237 AB622435 0.0148 0.0088 0.0088 0.0029 0.0000 0.0237 MN509083 0.0148 0.0088 0.0088 0.0029 0.0000 0.0237 0.0000 AB455803 0.0148 0.0088 0.0088 0.0029 0.0000 0.0237 0.0000 0.0000 Results of amino acid variation site analysis Amino acid variation site analysis was performed between 2024FXYQ153 and GI.6 in the reference sequence. The results showed that there were 4 amino acid site substitutions between 2024FXYQ153 and the VP1 partial region of the SaV GI.6 reference sequence. See Figure 2 for details. Discussion SaV and NoV are important pathogens that cause acute gastroenteritis in humans and can infect individuals of all age groups, with children being the most commonly infected population [5] . Globally, SaV infections account for approximately 10% of diarrhea cases and are estimated to cause 23,000 deaths annually among children under 5 years of age [6] . The clinical symptoms caused by SaV are milder than those caused by NoV. Therefore, there are more studies on NoV outbreaks at home and abroad. The detection methods for SaV are not yet widespread, and there are few reports on SaV outbreaks and studies on its molecular epidemiology. During this epidemic, the sick students were concentrated in the same class. Based on the results of hygienic investigations, epidemiological investigations, and laboratory tests, food poisoning was excluded, and it was inferred that this epidemic was a clustered epidemic of sapovirus infection transmitted through contact. SaV was first discovered in 1976 by electron microscopy (EM) of human diarrhea samples [7] . SaV can undergo recombination between the non-structural protein-encoding region (including the RdRp region) and the VP1 gene, resulting in new chimeric viruses [8] . The VP1 gene is more diverse than the RdRp gene and is an important region for genotyping [1] , and the VP1 protein is solely responsible for most capsid-related processes such as assembly, host interactions, and immunogenicity [9] . SaV has genotype diversity. in recent years, the genes of genotype GI are dominant globally and followed by genotype GII [10] GI.1 and GI.2 were dominant in genotype GI in Japan [11] , Canada [12], and in other countries [13] . Additionally, GI.1 is the most common in outbreaks and patients [14-16] . But Genotype GI.6 is rarely detected.In our study, the genotype of SaV causing gastroenteritis outbreaks in Nanchang in 2024 was GI.6. In China, there are few reports of SaV GI.6 outbreaks. In 2022 in Shenyang [2] ,2019-2023 in Beijing [17] , and 2023 in Zhejiang [18] , SaV GI.6 outbreaks were reported in universities, communities, kindergartens, and primary schools, respectively. Especially in Zhejiang Province between 2021 and 2023, GI.6 accounted for 57.14% of all genotypes in SaV outbreaks [18] . SaV GI.6 was also detected in scattered patients in Shandong, China [19], etc. Moreover, it was also detected in sewage in Africa [20] and Italy [21] . This indicates that the genotype may be widely prevalent in a large area. In our study, the nucleotide sequence homology of the capsid region genes of the 5 detected GI.6 strains in this study was 100%. The nucleotide homology between the partial VP1 region of the detected strains and the reference sequence of SaV GI.6 ranged from 97.07% to 99.12%, and the amino acid homology ranged from 96.49% to 97.37%. Whether there are mutations in the genes of other regions requires further whole-genome sequencing. In countries where the rotavirus vaccination programs have been implemented, the incidence of Sapovirus has increased, making it the third most common viral pathogen causing diarrhea, except rotavirus and norovirus, in children under 5 years old [22-23] . Since humans can produce short-term genotype-specific protective antibodies after being infected with SaV, and there is no immune protection against other genotypes [24] , and the SaV gene has high variability, especially in the regions other than the VP1 region that are related to the severity of infection. Therefore, in the future, it is necessary to further strengthen the whole-genome sequencing of the SaV and the analysis of mutation sites, and continuously monitor the recombination of the virus. This is crucial for understanding its genetic characteristics and controlling the outbreaks of acute gastroenteritis. Conclusion This epidemic is a viral gastroenteritis transmitted through contact that occurred in a primary school. The pathogen is SaV GI.6. Through phylogenetic tree and homology analysis, the genetic relationships and variation of the SaV GI.6 VP1 gene in our province were preliminarily revealed, filling the research gap in this field in our province and providing sequence references for the study of the molecular evolution of SaV infections in China. In future surveillance work, in-depth research on the molecular epidemiological characteristics of SaV with more samples should be continuously carried out to reveal all the genotypes, variations, and change patterns of the prevalent SaV in our province, so as to provide a scientific basis for better prevention and control of infant diarrhea. Declarations Ethics approval and consent to participate The study was approved by the Medical Ethics Committee of Jiangxi Provincial Center for Disease Control and Prevention with file number JJS-1.1/2024.12.31.And informed consent was obtained from all subjects and their legal guardians.All methods were performed in accordance with the relevant guidelines. Consent for publication Not applicable. Availability of data and materials The datasets generated and analysed during the current study are available in the National Center for Biotechnology Information repository,the genebank numbers are PX480016-PX480020.(The scheduled release date for our submission is Nov 4.2025) Competing interests The authors declare no competing interests. Funding Fund Project: Key Project of Science and Technology Innovation of Health Commission of Jiangxi Province, No.:2025ZD008. Authors' contributions Qian Wang and Shiwen Liu drafted the manuscript. Jun Zhou,Yanni Zhang,Xiaoqing Liu,,Fang Xiao and Yangbowen Wu were involved with sample processing, virus detection and VP1 amplification. Yuan xie,Gang Xu,Jiabo Du,Shuting Wei and Jianxiong Li were involved in data analysis. LG, LS and HM designed the project.Yong Shi,Dajin Xiao and Jianxiong Li provided important guidance. All authors reviewed and revised the first and final manuscript drafts. All authors read and agreed to the final manuscript. Acknowledgements We acknowledge support from the Jiangxi Provincial Key Laboratory of Major Epidemics Prevention and Control. References Oka T, Wang Q, Katayama K, Saif LJ. Comprehensive review of human sapoviruses. Clin Microbiol Rev. 2015;28(1):32-53. Tian J, Cong S, Chen L, et al.Molecular etiological study on the GI.6 sapovirus causing an infectious diarrhea outbreak in Shenyang, China. Chin J Epidemiol. 2023;44(8):1294-1300. Kumthip K, Khamrin P, Ushijima H, Maneekarn N. 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A confirmation of sapovirus re-infection gastroenteritis cases with different genogroups and genetic shifts in the evolving sapovirus genotypes, 2002-2011 [published correction appears in Arch Virol.2013 Dec;158(12):2641-2]. Arch Virol. 2012;157(10):1999-2003. Additional Declarations No competing interests reported. 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1","display":"","copyAsset":false,"role":"figure","size":214944,"visible":true,"origin":"","legend":"\u003cp\u003ePhylogenetic tree of nucleotide sequences in the partial capsid protein region of SaV\u003c/p\u003e\n\u003cp\u003eNote: The sequences marked with“▲” were obtained in this study.\u003c/p\u003e\n\u003cp\u003e\u003cbr\u003e\u003c/p\u003e","description":"","filename":"floatimage1.png","url":"https://assets-eu.researchsquare.com/files/rs-7881481/v1/7af82a00946ef81b1a8c9adb.png"},{"id":97119697,"identity":"4238926d-c2bd-4425-9a3e-db595a0727b9","added_by":"auto","created_at":"2025-12-01 07:47:03","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":329265,"visible":true,"origin":"","legend":"\u003cp\u003eAmino acid variation sites of the VP1 gene in SaV sequences and reference sequences in this study\u003c/p\u003e","description":"","filename":"floatimage2.png","url":"https://assets-eu.researchsquare.com/files/rs-7881481/v1/dae2af13a29a116c822e08d1.png"},{"id":97249440,"identity":"96a0daa7-900a-46a9-a820-d8ed9733df3f","added_by":"auto","created_at":"2025-12-02 13:12:36","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1258078,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-7881481/v1/10e52cae-f840-46dc-ae2b-95a8ffb113ff.pdf"},{"id":97119708,"identity":"dc6e9aa7-0814-4e8b-9872-cfd2718dc397","added_by":"auto","created_at":"2025-12-01 07:47:04","extension":"doc","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":113664,"visible":true,"origin":"","legend":"","description":"","filename":"SupplementaryMaterial.doc","url":"https://assets-eu.researchsquare.com/files/rs-7881481/v1/ec78fe235fafa143ece17f9e.doc"}],"financialInterests":"No competing interests reported.","formattedTitle":"Molecular Etiology and Epidemiological Features of a Sapovirus GI.6 Diarrheal Disease Cluster in Jiangxi, China","fulltext":[{"header":"Introduction","content":"\u003cp\u003eSapovirus (SaV) belongs to the genus \u003cem\u003eSapovirus\u003c/em\u003e in the family \u003cem\u003eCaliciviridae\u003c/em\u003e and is one of the important pathogens that cause human acute gastroenteritis (AGE)\u003csup\u003e[\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e]\u003c/sup\u003e. SaV is a single-stranded positive-sense RNA virus with a genome of approximately 7.1\u0026ndash;7.6 kb, and is divided into 2\u0026ndash;3 open reading frames (ORFs). ORF1 encodes six non-structural proteins NSP1\u0026thinsp;~\u0026thinsp;NSP7 and one major capsid structural protein VP1. ORF2 encodes the smaller structural protein VP2. The function of ORF3 remains unclear\u003csup\u003e[\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e]\u003c/sup\u003e. According to the nucleotide sequence differences of the capsid protein VP1 encoded by ORF1, SaV can be divided into 20 genotypes (GI-GXX), among which GI, GII, GIV, and GV can infect humans\u003csup\u003e[\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e]\u003c/sup\u003e. Human SaV is mainly transmitted through the fecal-oral route. Its clinical symptoms include dysphoria, nausea, vomiting, diarrhea, abdominal pain, and abdominal colic. According to sentinel surveillance data at both China national and Jiangxi provincial levels, sporadic cases occur throughout the year, with peak incidence observed during winter and spring seasons. It is characterized by diverse transmission routes, a low infectious dose, and general susceptibility of the population. Gastroenteritis induced by SaV is usually milder than that caused by NoV and is relatively rare in Jiangxi Province. However, with the widespread use of the rotavirus vaccine, outbreaks of acute gastroenteritis (AGE) caused by SaV in kindergartens, nursing homes, and schools are increasingly becoming a major public health issue\u003csup\u003e[\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e]\u003c/sup\u003e. In November 2024, a clustered outbreak of acute gastroenteritis occurred in a primary school in Nanchang, Jiangxi. This study conducted pathogen identification for this outbreak, as well as sequencing typing and genetic evolution analysis, aiming to understand the evolutionary characteristics of SaV and provide more effective strategies and means for the prevention and control of viral diarrhea.\u003c/p\u003e"},{"header":"Methods","content":"\u003cp\u003e\u003cstrong\u003eSample collection\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAccording to the definitions of suspected cases, clinically diagnosed cases, and confirmed cases in the \u0026quot;National Surveillance Program for Viral Diarrhea (2021 Edition)\u0026quot;, a suspected case is defined as any teacher, student, or staff member at the Yuquandao Campus of Nanchang Normal University Affiliated Primary School who has presented with gastrointestinal symptoms such as nausea, vomiting, abdominal pain, and diarrhea since November 5, 2024. A clinically diagnosed case refers to a suspected case that meets the criteria of having \u0026ge;3 bowel movements with a change in stool consistency (watery stools) within 24 hours and/or \u0026ge;2 episodes of vomiting within 24 hours, without other etiological diagnoses, and having an epidemiological association with a confirmed case. A confirmed case is a suspected or clinically diagnosed case with a positive nucleic acid test result for Sapovirus in fecal, anal swab, or vomitus samples. The Centers for Disease Control and Prevention (CDC) in Qingshanhu District conducted a case search by reviewing the school\u0026apos;s morning check records and records of absences due to illness, and interviewing school administrators and class teachers. On November 12, 2024, a total of 10 anal swab samples were collected from suspected cases on-site and sent to the district CDC laboratory for testing.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eSample processing and viral nucleic acid extraction\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAfter vortexing the collected anal swab samples at 2500 r/min for 2 minutes, let them stand for 5 minutes for later use. A fully automatic nucleic acid extractor (Strean SP96, Guangzhou Daan Gene) and its matching nucleic acid extraction or purification kit (catalog no. DA0602) were used to extract viral nucleic acid according to the kit instructions. The extracted nucleic acid was stored at 4\u0026deg;C for short-term storage and at -80\u0026deg;C for long-term storage.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eReal-time fluorescent RT-PCR (real-time RT-PCR)\u003c/strong\u003e\u003cstrong\u003e\u003cem\u003e\u0026nbsp;\u003c/em\u003e\u003c/strong\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eUsing the extracted viral nucleic acid as a template, perform real-time fluorescent RT-PCR detection of the virus with triple Real-time Fluorescent PCR Detection Kit for Group A Rotavirus/Adenovirus/Astrovirus Nucleic Acid (Beijing Applied Biological Technologies Co., Ltd. Catalog No.: A7133YH-50T) and Triple Real-time Fluorescent PCR Detection Kit for Norovirus Genogroup I/Norovirus Genogroup II/Sapovirus Nucleic Acid (Beijing Applied Biological Technologies Co., Ltd. Catalog No.: A7123YH-50T). The total reaction volume is 25 \u0026mu;L. The amplification conditions and result judgment shall be carried out according to the kit instructions. \u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAmplification and sequencing of the SaV capsid region (VP1)\u003c/strong\u003e\u003cstrong\u003e\u003cem\u003e\u0026nbsp;\u0026nbsp;\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eFor the samples positive for SaV in 1.3, the primer pair SLV5317 and SLV5749 was used to amplify the SaV VP1 gene [11]. SLV5317: 5\u0026rsquo;-CTC GCC ACC TAC RAW GCB TGG TT-3\u0026rsquo;, SLV5749: 5\u0026rsquo;-CGG RCY TCA AAV STA CCB CCC CA-3\u0026rsquo;. The reactions were performed using the One-Step RT-PCR Kit (QIAGEN, Germany; Cat. No. 210212) on an Eppendorf thermal cycler. The reaction system was as follows: 8 \u0026mu;L of 5xRT-PCR Buffer, 1.6 \u0026mu;L of dNTP mixture (10 mM) , 1.6 \u0026mu;L of RT-PCR Enzyme mixture,0.8 \u0026mu;L of SLV5317 (30 \u0026mu;M),0.8 \u0026mu;L of SLV5749 (30 \u0026mu;M),12 \u0026mu;L of DEPC-treated H₂O,and 8 \u0026mu;L of RNA template, for a total volume of 40 \u0026mu;L. The PCR conditions were as follows: reverse transcription at 50\u0026deg;C for 30 min; initial denaturation at 95\u0026deg;C for 15 min; 40 cycles of denaturation at 94\u0026deg;C for 30 s, annealing at 50\u0026deg;C for 30 s and extension at 72\u0026deg;C for 1 min, and a final extension at 72\u0026deg;C for 10 min; 4\u0026deg;C indefinitely. The expected PCR product size was 434bp. The PCR products were sent to Sangon Biotechnology (Shanghai) Co., Ltd. for sequencing.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eSequence splicing and molecular genetic evolution analysis of SaV VP1 \u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe SeqMan software was used to organize and splice the sequences. At the same time, the sequences of the VP1 region of SaV GI~GVII types were downloaded from GenBank. Using the ClustalW method in the MEGA 12.0 software, the sequences that had been sequenced and spliced were compared with the nucleotide sequences of the VP1 region of SaV downloaded from the GenBank database.The neighbor-joining algorithm and maximum composite likelihood model with 1000 bootstraps method was used to construct a phylogenetic tree. NCBI BLAST was used to analyze the nucleotide and amino acid sequence homology, and Megalin software was used to analyze the average genetic distance of the sequences and the amino acid variation sites. \u0026nbsp;\u0026nbsp;\u003c/p\u003e"},{"header":"Results","content":"\u003cp\u003e\u003cstrong\u003eBasic situation of the epidemic\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe primary school where the epidemic occurred has a total of 6 grades, including 64 classes, with a total of 2,890 students and 201 teaching and administrative staff. As of 15:00 on November 20, 2024, a total of 10 suspected cases were found, all of whom were teachers and students in Class 10, Grade 5. The male-to-female ratio of the cases was 2.33:1. Nine cases were students, and 1 case was a teacher. Combining the results of epidemiological investigations and laboratory tests, among them, 5 cases were confirmed cases, and 5 cases were clinically diagnosed cases. The main clinical manifestations of these 10 cases were vomiting, nausea, abdominal pain, and diarrhea, and 2 cases had dysphoria. The earliest onset time of the cases was 11:00 on November 8, and the latest onset time was 9:00 on November 12. The peak onset time was concentrated from 00:00 on November 11 to 12:00 on November 12.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eTable 1: Statistical table of clinical symptoms\u0026nbsp;\u003c/p\u003e\n\u003cdiv align=\"\"\u003e\n \u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 114px;\"\u003e\n \u003cp\u003eSymptoms\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 161px;\"\u003e\n \u003cp\u003eNumber of cases (N=10)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 119px;\"\u003e\n \u003cp\u003ePercentage (%)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 114px;\"\u003e\n \u003cp\u003eVomiting\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 161px;\"\u003e\n \u003cp\u003e8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 119px;\"\u003e\n \u003cp\u003e80.00\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 114px;\"\u003e\n \u003cp\u003eAbdominal pain\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 161px;\"\u003e\n \u003cp\u003e8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 119px;\"\u003e\n \u003cp\u003e80.00\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 114px;\"\u003e\n \u003cp\u003eNausea\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 161px;\"\u003e\n \u003cp\u003e6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 119px;\"\u003e\n \u003cp\u003e60.00\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 114px;\"\u003e\n \u003cp\u003eDiarrhea\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 161px;\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 119px;\"\u003e\n \u003cp\u003e30.00\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 114px;\"\u003e\n \u003cp\u003eDysphoria\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 161px;\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 119px;\"\u003e\n \u003cp\u003e20.00\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n\u003c/div\u003e\n\u003cp\u003e\u003cstrong\u003eReal-time RT-PCR detection results\u003c/strong\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAmong the 10 samples (all were anal swabs from 10 suspected cases), 5 cases were detected positive for SaV, with a positive rate of 50% (5/10), and the others were all negative. \u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eSequencing and analysis of the SaV VP1\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAll five positive samples detected in Section 3.2 exhibited successful amplification via RT-PCR, with the resulting amplicons matching the predicted fragment length. Forty-five reference sequences of the VP1 region of each typical and representative SaV genotype globally were selected in this study. All the reference strain sequences of each SaV genotype were from the National Center for Biotechnology Information (NCBI). Genetic evolutionary analysis was performed with the 5 sequences in this epidemic cluster, and a phylogenetic tree was constructed. The evolutionary analysis showed that the SaV in this outbreak belonged to SaV GI.6, which was most closely related to the isolate from Zhejiang, China in 2023 (OR351089). The phylogenetic tree is shown in Figure 1. \u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eResults of gene nucleotide and amino acid homology analysis\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eSince the sequencing results of the VP1 region of 5 SaV samples in this epidemic showed 100% homology after comparison, one of them (2024FXYQ153) was selected for nucleotide homology analysis with GI.6 in the reference sequence. The results showed that the nucleotide homology between 2024FXYQ153 and the VP1 partial region of the SaV GI.6 reference sequence ranged from 97.07% to 99.12%, and the amino acid homology ranged from 96.49% to 97.37%. See Table 2 for details.\u003c/p\u003e\n\u003cp\u003eTable 2: Results of nucleotide and amino acid homology analysis of the SaV sequence and the reference sequence of the VP1 gene in this study\u003c/p\u003e\n\u003cdiv align=\"\"\u003e\n \u003ctable border=\"0\" cellspacing=\"0\" cellpadding=\"0\" width=\"493\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 166px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u003cem\u003eSample ID/Genbank ID\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 159px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u003cem\u003eNucleotide homology\u003c/em\u003e\u003c/strong\u003e\u003cstrong\u003e\u003cem\u003e（\u003c/em\u003e\u003c/strong\u003e\u003cstrong\u003e\u003cem\u003e%\u003c/em\u003e\u003c/strong\u003e\u003cstrong\u003e\u003cem\u003e）\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 168px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u003cem\u003eAmino acid homology\u003c/em\u003e\u003c/strong\u003e\u003cstrong\u003e\u003cem\u003e（\u003c/em\u003e\u003c/strong\u003e\u003cstrong\u003e\u003cem\u003e%\u003c/em\u003e\u003c/strong\u003e\u003cstrong\u003e\u003cem\u003e）\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 166px;\"\u003e\n \u003cp\u003e2024FXYQ153\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 159px;\"\u003e\n \u003cp\u003e100.00\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 168px;\"\u003e\n \u003cp\u003e100.00\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 166px;\"\u003e\n \u003cp\u003eOR351089/ZJ/CN/2023\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 159px;\"\u003e\n \u003cp\u003e99.12\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 168px;\"\u003e\n \u003cp\u003e97.37\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 166px;\"\u003e\n \u003cp\u003eAB455803/JPN/2005\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 159px;\"\u003e\n \u003cp\u003e98.83\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 168px;\"\u003e\n \u003cp\u003e96.49\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 166px;\"\u003e\n \u003cp\u003eMN509083/US/2016\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 159px;\"\u003e\n \u003cp\u003e98.83\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 168px;\"\u003e\n \u003cp\u003e96.49\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 166px;\"\u003e\n \u003cp\u003eAB6224354/JPN/2008\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 159px;\"\u003e\n \u003cp\u003e98.83\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 168px;\"\u003e\n \u003cp\u003e96.49\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 166px;\"\u003e\n \u003cp\u003eLC380411/JPN/2005\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 159px;\"\u003e\n \u003cp\u003e98.83\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 168px;\"\u003e\n \u003cp\u003e96.49\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 166px;\"\u003e\n \u003cp\u003eMG012443/US/2015\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 159px;\"\u003e\n \u003cp\u003e98.53\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 168px;\"\u003e\n \u003cp\u003e96.49\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 166px;\"\u003e\n \u003cp\u003eOQ835589/SY/CN/2021\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 159px;\"\u003e\n \u003cp\u003e98.53\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 168px;\"\u003e\n \u003cp\u003e97.37\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 166px;\"\u003e\n \u003cp\u003eAJ606694/UK/2000\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 159px;\"\u003e\n \u003cp\u003e97.07\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 168px;\"\u003e\n \u003cp\u003e97.37\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n\u003c/div\u003e\n\u003cp\u003e\u003cstrong\u003eResults of average genetic distance analysis\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe average nucleotide genetic distance between 2024FXYQ153 and GI.6 in the reference sequences was analyzed. The results showed that the average genetic distance between 2024FXYQ153 and the partial VP1 region of the SaV GI.6 reference sequences ranged from 0 to 0.0330. See Table 3 for details.\u003c/p\u003e\n\u003cp\u003eTable 3 \u0026nbsp;Analysis of the average nucleotide genetic distance of the VP1 gene between the SaV sequences in this study and the reference sequences\u003c/p\u003e\n\u003cdiv align=\"\"\u003e\n \u003ctable border=\"0\" cellspacing=\"0\" cellpadding=\"0\" width=\"555\" class=\"fr-table-selection-hover\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"2\" style=\"width: 99px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u003cem\u003eSample ID/Genbank ID\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"9\" style=\"width: 456px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u003cem\u003eSample ID/Genbank ID\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 65px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e2024FXYQ153\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 51px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eOQ835589\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 47px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eOR351089\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eMG012443\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 50px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eLC380411\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 48px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eAJ606694\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 49px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eAB622435\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 48px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eMN509083\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 46px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eAB455803\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 99px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e2024FXYQ153\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 65px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 51px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 47px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 50px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 48px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 49px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 48px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 46px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 99px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eOQ835589\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 65px;\"\u003e\n \u003cp\u003e0.0178\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 51px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 47px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 50px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 48px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 49px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 48px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 46px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 99px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eOR351089\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 65px;\"\u003e\n \u003cp\u003e0.0118\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 51px;\"\u003e\n \u003cp\u003e0.0177\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 47px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 50px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 48px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 49px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 48px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 46px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 99px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eMG012443\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 65px;\"\u003e\n \u003cp\u003e0.0178\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 51px;\"\u003e\n \u003cp\u003e0.0118\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 47px;\"\u003e\n \u003cp\u003e0.0117\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 50px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 48px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 49px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 48px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 46px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 99px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eLC380411\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 65px;\"\u003e\n \u003cp\u003e0.0148\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 51px;\"\u003e\n \u003cp\u003e0.0088\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 47px;\"\u003e\n \u003cp\u003e0.0088\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003e0.0029\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 50px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 48px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 49px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 48px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 46px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 99px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eAJ606694\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 65px;\"\u003e\n \u003cp\u003e0.0330\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 51px;\"\u003e\n \u003cp\u003e0.0329\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 47px;\"\u003e\n \u003cp\u003e0.0298\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003e0.0267\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 50px;\"\u003e\n \u003cp\u003e0.0237\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 48px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 49px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 48px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 46px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 99px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eAB622435\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 65px;\"\u003e\n \u003cp\u003e0.0148\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 51px;\"\u003e\n 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\u003cp\u003e\u003cstrong\u003eAB455803\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 65px;\"\u003e\n \u003cp\u003e0.0148\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 51px;\"\u003e\n \u003cp\u003e0.0088\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 47px;\"\u003e\n \u003cp\u003e0.0088\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003e0.0029\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 50px;\"\u003e\n \u003cp\u003e0.0000\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 48px;\"\u003e\n \u003cp\u003e0.0237\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 49px;\"\u003e\n \u003cp\u003e0.0000\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 48px;\"\u003e\n \u003cp\u003e0.0000\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 46px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n\u003c/div\u003e\n\u003cp\u003e\u003cstrong\u003eResults of amino acid variation site analysis\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAmino acid variation site analysis was performed between 2024FXYQ153 and GI.6 in the reference sequence. The results showed that there were 4 amino acid site substitutions between 2024FXYQ153 and the VP1 partial region of the SaV GI.6 reference sequence. See Figure 2 for details.\u0026nbsp;\u003c/p\u003e"},{"header":"Discussion","content":"\u003cp\u003eSaV and NoV are important pathogens that cause acute gastroenteritis in humans and can infect individuals of all age groups, with children being the most commonly infected population\u003csup\u003e[5]\u003c/sup\u003e. Globally, SaV infections account for approximately 10% of diarrhea cases and are estimated to cause 23,000 deaths annually among children under 5 years of age\u003csup\u003e[6]\u003c/sup\u003e. The clinical symptoms caused by SaV are milder than those caused by NoV. Therefore, there are more studies on NoV outbreaks at home and abroad. The detection methods for SaV are not yet widespread, and there are few reports on SaV outbreaks and studies on its molecular epidemiology.\u003c/p\u003e\n\u003cp\u003eDuring this epidemic, the sick students were concentrated in the same class. Based on the results of hygienic investigations, epidemiological investigations, and laboratory tests, food poisoning was excluded, and it was inferred that this epidemic was a clustered epidemic of sapovirus infection transmitted through contact.\u003c/p\u003e\n\u003cp\u003eSaV was first discovered in 1976 by electron microscopy (EM) of human diarrhea samples\u003csup\u003e[7]\u003c/sup\u003e. SaV can undergo recombination between the non-structural protein-encoding region (including the RdRp region) and the VP1 gene, resulting in new chimeric viruses\u003csup\u003e[8]\u003c/sup\u003e. The VP1 gene is more diverse than the RdRp gene and is an important region for genotyping\u003csup\u003e[1]\u003c/sup\u003e, and the VP1 protein is solely responsible for most capsid-related processes such as assembly, host interactions, and immunogenicity\u003csup\u003e[9]\u003c/sup\u003e. SaV has genotype diversity. in recent years, the genes of genotype GI are dominant globally and followed by genotype GII\u003csup\u003e[10]\u003c/sup\u003e GI.1 and GI.2 were dominant in genotype GI in Japan\u003csup\u003e[11]\u003c/sup\u003e, Canada\u003csup\u003e[12],\u0026nbsp;\u003c/sup\u003eand in other countries\u003csup\u003e[13]\u003c/sup\u003e. Additionally, GI.1 is the most common in outbreaks and patients\u003csup\u003e[14-16]\u003c/sup\u003e. But Genotype GI.6 is rarely detected.In our study, the genotype of SaV causing gastroenteritis outbreaks in Nanchang in 2024 was GI.6. In China, there are few reports of SaV GI.6 outbreaks. In 2022 in Shenyang\u003csup\u003e[2]\u003c/sup\u003e,2019-2023 in Beijing\u003csup\u003e[17]\u003c/sup\u003e, and 2023 in Zhejiang\u003csup\u003e[18]\u003c/sup\u003e, SaV GI.6 outbreaks were reported in universities, communities, kindergartens, and primary schools, respectively. Especially in Zhejiang Province between 2021 and 2023, GI.6 accounted for 57.14% of all genotypes in SaV outbreaks\u003csup\u003e[18]\u003c/sup\u003e. SaV GI.6 was also detected in scattered patients in Shandong, China\u003csup\u003e[19],\u0026nbsp;\u003c/sup\u003eetc. Moreover, it was also detected in sewage in Africa\u003csup\u003e[20]\u003c/sup\u003eand Italy\u003csup\u003e[21]\u003c/sup\u003e. This indicates that the genotype may be widely prevalent in a large area. In our study, the nucleotide sequence homology of the capsid region genes of the 5 detected GI.6 strains in this study was 100%. The nucleotide homology between the partial VP1 region of the detected strains and the reference sequence of SaV GI.6 ranged from 97.07% to 99.12%, and the amino acid homology ranged from 96.49% to 97.37%. Whether there are mutations in the genes of other regions requires further whole-genome sequencing.\u003c/p\u003e\n\u003cp\u003eIn countries where the rotavirus vaccination programs have been implemented, the incidence of Sapovirus has increased, making it the third most common viral pathogen causing diarrhea, except rotavirus and norovirus, in children under 5 years old \u003csup\u003e[22-23]\u003c/sup\u003e. Since humans can produce short-term genotype-specific protective antibodies after being infected with SaV, and there is no immune protection against other genotypes\u003csup\u003e[24]\u003c/sup\u003e, and the SaV gene has high variability, especially in the regions other than the VP1 region that are related to the severity of infection. Therefore, in the future, it is necessary to further strengthen the whole-genome sequencing of the SaV and the analysis of mutation sites, and continuously monitor the recombination of the virus. This is crucial for understanding its genetic characteristics and controlling the outbreaks of acute gastroenteritis. \u0026nbsp;\u003c/p\u003e"},{"header":"Conclusion","content":"\u003cp\u003eThis epidemic is a viral gastroenteritis transmitted through contact that occurred in a primary school. The pathogen is SaV GI.6. Through phylogenetic tree and homology analysis, the genetic relationships and variation of the SaV GI.6 VP1 gene in our province were preliminarily revealed, filling the research gap in this field in our province and providing sequence references for the study of the molecular evolution of SaV infections in China. In future surveillance work, in-depth research on the molecular epidemiological characteristics of SaV with more samples should be continuously carried out to reveal all the genotypes, variations, and change patterns of the prevalent SaV in our province, so as to provide a scientific basis for better prevention and control of infant diarrhea. \u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cbr\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eEthics approval and consent to participate\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe study was approved by the Medical Ethics Committee of Jiangxi Provincial Center for Disease Control and Prevention with file number JJS-1.1/2024.12.31.And informed consent was obtained from all subjects and their legal guardians.All methods were performed in accordance with the relevant guidelines.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent for publication\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAvailability of data and materials\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe datasets generated and analysed during the current study are available in the \u003cem\u003eNational Center for Biotechnology Information\u003c/em\u003e repository,the genebank numbers are PX480016-PX480020.(The scheduled release date for our submission is Nov 4.2025)\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCompeting interests\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors declare no competing interests.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eFund Project: Key Project of Science and Technology Innovation of Health Commission of Jiangxi Province, No.:2025ZD008.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthors\u0026apos; contributions\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eQian Wang and Shiwen Liu drafted the manuscript. Jun Zhou,Yanni Zhang,Xiaoqing Liu,,Fang Xiao and Yangbowen Wu were involved with sample processing, virus detection and VP1 amplification. Yuan xie,Gang Xu,Jiabo Du,Shuting Wei and Jianxiong Li were involved in data analysis. LG, LS and HM designed the project.Yong Shi,Dajin Xiao and Jianxiong Li provided important guidance. All authors reviewed and revised the first and final manuscript drafts. All authors read and agreed to the final manuscript.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAcknowledgements\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eWe acknowledge support from the Jiangxi Provincial Key Laboratory of Major Epidemics Prevention and Control.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eOka T, Wang Q, Katayama K, Saif LJ. Comprehensive review of human sapoviruses. Clin Microbiol Rev. 2015;28(1):32-53.\u003c/li\u003e\n\u003cli\u003eTian J, Cong S, Chen L, et al.Molecular etiological study on the GI.6 sapovirus causing an infectious diarrhea outbreak in Shenyang, China. Chin J Epidemiol. 2023;44(8):1294-1300.\u003c/li\u003e\n\u003cli\u003eKumthip K, Khamrin P, Ushijima H, Maneekarn N. Enteric and non-enteric adenoviruses associated with acute gastroenteritis in pediatric patients in Thailand, 2011 to 2017. PLoS One. 2019;14(8):e0220263. \u003c/li\u003e\n\u003cli\u003eRouhani S, Pe\u0026ntilde;ataro Yori P, Paredes Olortegui M, et al. The Epidemiology of Sapovirus in the Etiology, Risk Factors, and Interactions of Enteric Infection and Malnutrition and the Consequences for Child Health and Development Study: Evidence of Protection Following Natural Infection. Clin Infect Dis. 2022;75(8):1334-1341.\u003c/li\u003e\n\u003cli\u003eBecker-Dreps S, Gonz\u0026aacute;lez F, Bucardo F. Sapovirus: an emerging cause of childhood diarrhea. Curr Opin Infect Dis. 2020;33(5):388-397.\u003c/li\u003e\n\u003cli\u003eCohen AL, Platts-Mills JA, Nakamura T, et al. Aetiology and incidence of diarrhoea requiring hospitalisation in children under 5 years of age in 28 low-income and middle-income countries: findings from the Global Pediatric Diarrhea Surveillance network. BMJ Glob Health. 2022;7(9):e009548.\u003c/li\u003e\n\u003cli\u003eMadeley, C.R.; Cosgrove, B.P. Letter: Caliciviruses in man. Lancet 1976, 1, 199-200\u003c/li\u003e\n\u003cli\u003eMizukoshi F, Kimura R, Shirai T, et al. Molecular Evolutionary Analyses of the RNA-Dependent RNA Polymerase (RdRp) Region and VP1 Gene in Sapovirus GI.1 and GI.2. Microorganisms. 2025;13(2):322.\u003c/li\u003e\n\u003cli\u003eMiyazaki N, Song C, Oka T, et al. Atomic Structure of the Human Sapovirus Capsid Reveals a Unique Capsid Protein Conformation in Caliciviruses. J Virol. 2022;96(9):e0029822.\u003c/li\u003e\n\u003cli\u003eRazizadeh MH, Khatami A, Zarei M. Global molecular prevalence and genotype distribution of Sapovirus in children with gastrointestinal complications: a systematic review and meta-analysis. Rev Med Virol. 2022;32(3):e2302.\u003c/li\u003e\n\u003cli\u003eDoan YH, Yamashita Y, Shinomiya H, Motoya T, Sakon N, Suzuki R, et al. Distribution of human sapovirus strain genotypes over the last four decades in Japan: a global perspective. Jpn J Infect Dis. 2023.\u003c/li\u003e\n\u003cli\u003eZhuo R, Ding X, Freedman SB, et al. Molecular Epidemiology of Human Sapovirus among Children with Acute Gastroenteritis in Western Canada. J Clin Microbiol. 2021;59(10):e0098621. \u003c/li\u003e\n\u003cli\u003eMizukoshi F, Kimura R, Shirai T, et al. Molecular Evolutionary Analyses of the RNA-Dependent RNA Polymerase (RdRp) Region and VP1 Gene in Sapovirus GI.1 and GI.2. Microorganisms. 2025;13(2):322.\u003c/li\u003e\n\u003cli\u003eJi X, Guo C, Dai Y, et al. Genomic Characterization and Molecular Evolution of Sapovirus in Children under 5 Years of Age. Viruses. 2024;16(1):146.\u003c/li\u003e\n\u003cli\u003eGonz\u0026aacute;lez F, Diez-Valcarce M, Reyes Y, et al. Timing and genotype distribution of symptomatic and asymptomatic sapovirus infections and re-infections in a Nicaraguan birth cohort. Clin Microbiol Infect. 2023;29(4):540.e9-540.e15.\u003c/li\u003e\n\u003cli\u003eHoque SA, Akari Y, Khamrin P, et al. Epidemiology and molecular evolution of GI.1 sapovirus in the recent era. J Med Virol. 2024;96(9):e29904.\u003c/li\u003e\n\u003cli\u003eJing H,Wei W, He M,et al.Epidemiological and genetic characteristics of sapovirus outbreaks in Beijing,2019-2023.Chin J Virol.2024;40(2):15-22.\u003c/li\u003e\n\u003cli\u003eSu L, Mao H, Sun Y, et al.Genotype analysis of Sapovirus outbreaks in Zhejiang Province, China. BMC Infect Dis. 2023;23:123.\u003c/li\u003e\n\u003cli\u003eGuo M, Li M, Liu T, et al. Epidemiological and Genetic Characteristics of Sapovirus in Shandong, China, 2022-2023. Viruses. 2025;17(4):469.\u003c/li\u003e\n\u003cli\u003eMakhaola K, Moyo S, Kebaabetswe LP. Distribution and Genetic Variability of Sapoviruses in Africa. Viruses. 2020;12(5):490.\u003c/li\u003e\n\u003cli\u003eMancini P, Bonanno Ferraro G, Iaconelli M, et al. Molecular characterization of human Sapovirus in untreated sewage in Italy by amplicon-based Sanger and next-generation sequencing. J Appl Microbiol. 2019;126(1):324-331\u003c/li\u003e\n\u003cli\u003ePitk\u0026auml;nen O, Vesikari T, Hemming-Harlo M. The role of the sapovirus infection increased in gastroenteritis after national immunisation was introduced. Acta Paediatr. 2019;108(7):1338-1344.\u003c/li\u003e\n\u003cli\u003eHalasa N, Piya B, Stewart LS, et al. The Changing Landscape of Pediatric Viral Enteropathogens in the Post-Rotavirus Vaccine Era. Clin Infect Dis. 2021;72(4):576-585.\u003c/li\u003e\n\u003cli\u003eHarada S, Oka T, Tokuoka E, et al. A confirmation of sapovirus re-infection gastroenteritis cases with different genogroups and genetic shifts in the evolving sapovirus genotypes, 2002-2011 [published correction appears in Arch Virol.2013 Dec;158(12):2641-2]. Arch Virol. 2012;157(10):1999-2003.\u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"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":"scientific-reports","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"scirep","sideBox":"Learn more about [Scientific Reports](http://www.nature.com/srep/)","snPcode":"","submissionUrl":"","title":"Scientific Reports","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Scientific Reports","inReviewEnabled":true,"inReviewRevisionsEnabled":true},"keywords":"Sapovirus, acute gastroenteritis, epidemic cluster, molecular epidemiology","lastPublishedDoi":"10.21203/rs.3.rs-7881481/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-7881481/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003ch2\u003eObjective\u003c/h2\u003e\u003cp\u003eTo clarify the etiological and genetic evolutionary characteristics of an acute gastroenteritis cluster epidemic caused by sapovirus in Nanchang, Jiangxi, China.\u003c/p\u003e\u003ch2\u003eMethods\u003c/h2\u003e\u003cp\u003eA total of 10 anal swab samples were collected from student and teacher cases during a gastroenteritis epidemic at a primary school in Nanchang, Jiangxi Province in November 2024. Real-time fluorescent RT-PCR was used for routine gene detection of diarrhea viruses. For the sapovirus nucleic acid-positive samples, traditional RT-PCR was used for amplification, sequence determination, splicing, and genetic evolutionary analysis.\u003c/p\u003e\u003ch2\u003eResults\u003c/h2\u003e\u003cp\u003eThis epidemic cluster was formed due to contact transmission. Real-time fluorescent RT-PCR detection confirmed that 5 samples were positive for sapovirus nucleic acid, and the capsid protein gene VP1 fragments were successfully amplified. Genetic evolutionary analysis revealed that the molecular type of sapovirus was the GI.6 genotype, with 100% homology among the 5 sequences. The nucleotide homology between the sequences in this study and the sapovirus GI.6 reference sequence ranged from 97.07% to 99.12%, and the amino acid homology ranged from 96.49% to 97.37%. Amino acid variation site analysis revealed that there were 4 amino acid site substitutions between them. The closest distance of nucleotide homology was 99.12% from Zhejiang, China in 2023 (OR351089)。\u003c/p\u003e\u003ch2\u003eConclusion\u003c/h2\u003e\u003cp\u003eThe causative agent of this epidemic cluster was identified as sapovirus GI.6, a genotype rarely reported in Jiangxi. In the future, further research on the molecular epidemiology of sapovirus with more gene sites that may mutate should be strengthened.\u003c/p\u003e","manuscriptTitle":"Molecular Etiology and Epidemiological Features of a Sapovirus GI.6 Diarrheal Disease Cluster in Jiangxi, China","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-12-01 07:46:57","doi":"10.21203/rs.3.rs-7881481/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2025-11-28T08:37:26+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-11-27T09:44:37+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"259445107491097781213268654548487910003","date":"2025-11-25T20:07:00+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-11-25T08:38:33+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"168869171564032181731686459595904174135","date":"2025-11-24T23:43:50+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"145068074205933234646242190558557945147","date":"2025-11-24T20:31:44+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"126939128627096563235669110319767623921","date":"2025-11-24T04:59:46+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2025-11-23T23:57:18+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2025-11-23T23:54:39+00:00","index":"","fulltext":""},{"type":"editorInvited","content":"","date":"2025-11-11T05:01:31+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2025-11-10T04:14:56+00:00","index":"","fulltext":""},{"type":"submitted","content":"Scientific Reports","date":"2025-11-10T04:12:13+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"scientific-reports","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"scirep","sideBox":"Learn more about [Scientific Reports](http://www.nature.com/srep/)","snPcode":"","submissionUrl":"","title":"Scientific Reports","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Scientific Reports","inReviewEnabled":true,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"532ccd86-2d34-4e12-9caa-00630ca4c64a","owner":[],"postedDate":"December 1st, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"under-review","subjectAreas":[{"id":58737725,"name":"Health sciences/Diseases"},{"id":58737726,"name":"Biological sciences/Genetics"},{"id":58737727,"name":"Biological sciences/Microbiology"},{"id":58737728,"name":"Biological sciences/Molecular biology"}],"tags":[],"updatedAt":"2026-05-20T01:23:30+00:00","versionOfRecord":[],"versionCreatedAt":"2025-12-01 07:46:57","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-7881481","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-7881481","identity":"rs-7881481","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}