中国动物检疫2025，Vol.42Issue(9)：69-76,8.DOI:10.3969/j.issn.1005-944X.2025.09.012
A型牛轮状病毒SYBR Green Ⅰ RT-qPCR检测方法的建立
Development of a SYBR Green Ⅰ RT-qPCR Assay for Bovine Rotavirus A
摘要
Abstract
Cattle industry is endangered by bovine rotavirus A(BRV-A)to the maximum extent,as the virus is one of the major pathogens causing bovine diarrhea.A SYBR Green Ⅰ RT-qPCR assay for detecting the virus was established through designing specific primers targeting the conserved region of its VP6 gene,which was expected to be rapid,high-throughput and cost-effective.The results showed that a good linear correlation was observed between the amplification Ct value of the plasmid standard with a concentration gradient of 4.41 × 108-4.41 × 102 copies/μL and copy number concentration,generating single-peak melting curves;there was no cross-reactivity with other common pathogens causing bovine diarrheal,the limit of detection(LOD)was 4.41 × 101 copies/μL,the coefficient of variation(CV)for both intra-and inter-assay reproducibility was＜1％;positive detection rate against fecal samples was higher than that by the PCR assay defined in local standards,and the test results were highly consistent.In conclusion,the BRV-A SYBR Green Ⅰ RT-qPCR assay established in the study was sensitive,specific,stable,accessible and cost-effective,technically supporting intensive detection of BRV-A from clinical samples and early diagnosis of relevant infection.关键词
牛轮状病毒/SYBR Green Ⅰ荧光RT-PCR/检测方法Key words
BRV/SYBR Green Ⅰ fluorescent RT-PCR/test method分类
农业科技引用本文复制引用
焦迪,李军燕,赵治国,王瑞,郝艳霞,张晓倩,崔强,王海艳,陈林军,延涵,罗晓平..A型牛轮状病毒SYBR Green Ⅰ RT-qPCR检测方法的建立[J].中国动物检疫,2025,42(9):69-76,8.基金项目
内蒙古自治区重点研发和成果转化计划项目(2023YFDZ0048) （2023YFDZ0048）
中央引导地方科技发展资金项目(2024ZY0156) （2024ZY0156）
海关总署科技计划项目(2023HK012,2024HK261) （2023HK012,2024HK261）
鄂尔多斯市国家可持续发展议程创新示范区建设(含厅市会商、科技突围工程)科技支撑项目(KCX2024016) （含厅市会商、科技突围工程）
内蒙古自治区科研创新团队建设专项B类团队(二类团队)项目(BR251303) （二类团队）
国家乳业技术创新中心项目(2023-JSGG-5) （2023-JSGG-5）
