实验动物与比较医学2026，Vol.46Issue(3)：408-415,8.DOI:10.12300/j.issn.1674-5817.2025.130
MHV、MPV和Reo-3三重TaqMan定量PCR检测方法的建立
Development of a Triplex TaqMan Quantitative PCR Method for Simultaneous Detection of MHV,MPV,and Reo-3
摘要
Abstract
Objective To establish a highly specific and sensitive triplex TaqMan quantitative PCR method for the detection of mouse hepatitis virus(MHV),mouse parvovirus(MPV),and reovirus type 3(Reo-3)in laboratory mice.Methods Recombinant plasmid standards were constructed using conserved genomic regions of the three target viruses.Specific primers and probes were designed,and a triplex TaqMan quantitative PCR(qPCR)system was optimized through preliminary experiments.Sensitivity was evaluated using 10-fold serial dilutions(101-107 copies/μL).Intra-and inter-assay repeatability were assessed via reproducibility experiments.Results A strong linear correlation(R 2>0.99)was observed between copy number and Ct value at template concentrations of 102-107 copies/μL.Intra-assay and inter-assay coefficients of variation(CV)were both below 5%.No cross-reactivity was observed with mouse cytomegalovirus(MCMV),vesicular stomatitis virus(VSV),Sendai virus(SeV),or pneumonia virus of mice(PVM).Conclusion The triplex TaqMan quantitative PCR method established in this study has high sensitivity,good repeatability,and strong specificity.It enables the rapid,specific,and sensitive detection of MHV,MPV,and Reo-3,allows the simultaneous detection of multiple pathogens in a single tube,and can be applied to the detection of clinical samples from laboratory animals and to epidemiological investigations,thereby providing effective scientific and technological support for the prevention and control of viral infection and the interruption of transmission.关键词
小鼠肝炎病毒/小鼠细小病毒/呼肠孤病毒Ⅲ型/三重TaqMan定量PCRKey words
Mouse hepatitis virus/Mouse parvovirus/Reovirus type 3/Triplex TaqMan quantitative PCR分类
生物科学引用本文复制引用
周忆旻,张昕宇,杨佳诺,刘梦佳,司灿灿,叶海绿,孙文超,兰添..MHV、MPV和Reo-3三重TaqMan定量PCR检测方法的建立[J].实验动物与比较医学,2026,46(3):408-415,8.基金项目
温州市基础性医疗卫生科技项目"基于数字PCR技术的SPF级实验动物病毒病原体快速检测关键技术研究与应用"(Y20220136) （Y20220136）
浙江省高校实验室工作研究项目"中外合作办学背景下高校SPF级动物实验室建设与管理机制初探—以温州肯恩大学为例"(YB202431) （YB202431）
