现代畜牧兽医Issue(3)：11-15,5.DOI:10.20154/j.cnki.issn1672-9692.2025.03.003
1种牛轮状病毒Taq-Man荧光定量RT-PCR检测方法的建立
Establishment of a Taq-Man fluorescent quantitative RT-PCR detection method for bovine rotavirus
王秀明 1陈君彦 2础鲁 3刘艳霞1
作者信息
- 1. 辽宁益康生物股份有限公司,辽宁 辽阳 111000
- 2. 金宇保灵生物药品有限公司,内蒙古 呼和浩特 010111
- 3. 阿拉善左旗农牧业综合行政执法大队,内蒙古 巴彦浩特 750300
- 折叠
摘要
Abstract
The experiment aimed to establish a simple and rapid Taq-Man fluorescent quantitative RT-PCR method for the detection of bovine rotavirus(BRV).By analyzing and comparing the BRV VP7 gene sequences downloaded from NCBI,two pairs of specific primers and a Taq-Man probe were designed in the conserved region of the VP7 gene.After optimizing the reaction conditions,a TaqMan fluorescent quantitative RT-PCR method for BRV was established.The established detection method was used to test the standard products,plot the standard curve,and evaluate the specificity,sensitivity,and stability of the detection method.Clinical samples and vaccine semi-finished products were also tested.The results showed that the lowest detection limit of the established Taq-Man fluorescent quantitative RT-PCR method was 10 copies/μL,and the intra-group and inter-group repeatability coefficients of variation(CV)were both less than 3.50%.The detection of clinical samples and vaccine semi-finished products showed that all vaccine semi-finished products were positive,and the positive rate of clinical samples was 69%.The study demonstrated that the BRV Taq-Man fluorescent quantitative RT-PCR method is simple to operate,highly specific,and sensitive,and has important reference value for the diagnosis and detection of BRV and vaccine production.关键词
牛轮状病毒(BRV)/Taq-Man探针/荧光定量RT-PCR/VP7基因Key words
Bovine rotavirus(BRV)/Taq-Man probe/Fluorescent quantitative RT-PCR/VP7 gene分类
农业科技引用本文复制引用
王秀明,陈君彦,础鲁,刘艳霞..1种牛轮状病毒Taq-Man荧光定量RT-PCR检测方法的建立[J].现代畜牧兽医,2025,(3):11-15,5.
