{"title":"开发轮状病毒 G3P8 和 G9P8 的 RT-dPCR 方法和参考材料。","authors":"Jiayi Yang, Mingwei Liu, Huijie Li, Yunhua Gao, Lianhua Dong","doi":"10.1007/s00216-024-05690-2","DOIUrl":null,"url":null,"abstract":"<p><p>The rotavirus (RV) is the predominant causative pathogen associated with acute gastroenteritis in children aged below 5 years, leading to an annual mortality rate of 200,000 infants globally. Despite the development of a vaccine, it exacerbates the medical burden around the world. Here, we have developed reverse transcription digital PCR (RT-dPCR) methods for precise and absolute quantification of nucleic acid in rotavirus G3P8 and G9P8. The pseudovirus reference material (RM) contained the RNA fragment encoding VP4 and VP7. The assigned values with expanded uncertainty were determined as (2432 ± 510) copies/μL and (3406 ± 613) copies/μL. The RM and RT-dPCR methods were employed to validate various digital platforms, revealing the inadequate performance of platform III, which could potentially result in \"false-negative\" outcomes. The application of RT-dPCR techniques and pseudovirus RM confers advantages in the diagnosis of RV-induced diseases, thereby enhancing the survival rate of young children suffering from acute gastroenteritis.</p>","PeriodicalId":462,"journal":{"name":"Analytical and Bioanalytical Chemistry","volume":" ","pages":""},"PeriodicalIF":3.8000,"publicationDate":"2024-12-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Development of RT-dPCR method and reference material for rotavirus G3P8 and G9P8.\",\"authors\":\"Jiayi Yang, Mingwei Liu, Huijie Li, Yunhua Gao, Lianhua Dong\",\"doi\":\"10.1007/s00216-024-05690-2\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>The rotavirus (RV) is the predominant causative pathogen associated with acute gastroenteritis in children aged below 5 years, leading to an annual mortality rate of 200,000 infants globally. Despite the development of a vaccine, it exacerbates the medical burden around the world. Here, we have developed reverse transcription digital PCR (RT-dPCR) methods for precise and absolute quantification of nucleic acid in rotavirus G3P8 and G9P8. The pseudovirus reference material (RM) contained the RNA fragment encoding VP4 and VP7. The assigned values with expanded uncertainty were determined as (2432 ± 510) copies/μL and (3406 ± 613) copies/μL. The RM and RT-dPCR methods were employed to validate various digital platforms, revealing the inadequate performance of platform III, which could potentially result in \\\"false-negative\\\" outcomes. The application of RT-dPCR techniques and pseudovirus RM confers advantages in the diagnosis of RV-induced diseases, thereby enhancing the survival rate of young children suffering from acute gastroenteritis.</p>\",\"PeriodicalId\":462,\"journal\":{\"name\":\"Analytical and Bioanalytical Chemistry\",\"volume\":\" \",\"pages\":\"\"},\"PeriodicalIF\":3.8000,\"publicationDate\":\"2024-12-16\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Analytical and Bioanalytical Chemistry\",\"FirstCategoryId\":\"92\",\"ListUrlMain\":\"https://doi.org/10.1007/s00216-024-05690-2\",\"RegionNum\":2,\"RegionCategory\":\"化学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q1\",\"JCRName\":\"BIOCHEMICAL RESEARCH METHODS\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Analytical and Bioanalytical Chemistry","FirstCategoryId":"92","ListUrlMain":"https://doi.org/10.1007/s00216-024-05690-2","RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"BIOCHEMICAL RESEARCH METHODS","Score":null,"Total":0}
Development of RT-dPCR method and reference material for rotavirus G3P8 and G9P8.
The rotavirus (RV) is the predominant causative pathogen associated with acute gastroenteritis in children aged below 5 years, leading to an annual mortality rate of 200,000 infants globally. Despite the development of a vaccine, it exacerbates the medical burden around the world. Here, we have developed reverse transcription digital PCR (RT-dPCR) methods for precise and absolute quantification of nucleic acid in rotavirus G3P8 and G9P8. The pseudovirus reference material (RM) contained the RNA fragment encoding VP4 and VP7. The assigned values with expanded uncertainty were determined as (2432 ± 510) copies/μL and (3406 ± 613) copies/μL. The RM and RT-dPCR methods were employed to validate various digital platforms, revealing the inadequate performance of platform III, which could potentially result in "false-negative" outcomes. The application of RT-dPCR techniques and pseudovirus RM confers advantages in the diagnosis of RV-induced diseases, thereby enhancing the survival rate of young children suffering from acute gastroenteritis.
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Analytical and Bioanalytical Chemistry’s mission is the rapid publication of excellent and high-impact research articles on fundamental and applied topics of analytical and bioanalytical measurement science. Its scope is broad, and ranges from novel measurement platforms and their characterization to multidisciplinary approaches that effectively address important scientific problems. The Editors encourage submissions presenting innovative analytical research in concept, instrumentation, methods, and/or applications, including: mass spectrometry, spectroscopy, and electroanalysis; advanced separations; analytical strategies in “-omics” and imaging, bioanalysis, and sampling; miniaturized devices, medical diagnostics, sensors; analytical characterization of nano- and biomaterials; chemometrics and advanced data analysis.
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