Establishment of a Taq-Man fluorescent quantitative RT-PCR detection method for bovine rotavirus
Wang Xiuming
Chen Junyan
Chu Lu
Liu Yanxia
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.
Keywords：Bovine rotavirus(BRV)Taq-Man probeFluorescent quantitative RT-PCRVP7 gene
Publication Date：2025-03-15
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：5( 11-15 )