中兽医医药杂志2026，Vol.45Issue(5)：71-76,6.DOI:10.13823/j.cnki.jtcvm.2026.050
啮齿类实验动物仙台病毒和小鼠肺炎病毒双重PCR检测方法的建立
Establishment of a duplex PCR detection method for Sendai virus and Pneumonia virus of mice in rodent laboratory animals
摘要
Abstract
Sendai virus(SeV)and Pneumonia virus of mice(PVM)are prevalent susceptible viruses in rodent laboratory animals.Infected animals predominantly display respiratory symptoms,which exert significant impacts on both the health of laboratory animals and the reliability of experimental results.This study aimed to establish a simple,rapid and efficient duplex PCR detection method for SeV and PVM,which can be used for pathogen monitoring of laboratory animals and related samples.Specific primers were designed based on the SeV L gene and PVM NS2 gene.A duplex PCR system was established by optimizing the concentration of primers,and testing the specificity and sensitivity of this system.After systematic optimization,0.40 μmol/L of upstream and downstream primers for PVM and SeV were determined,and the annealing temperature was 60℃.The results showed that the optimized duplex PCR assay could specifically amplify the SeV and PVM templates in mixed samples without cross reaction.Both quantitative real-time PCR(qPCR)and the duplex PCR could amplify the target viral DNA fragments from standard viral template,but no fragments were detected in the negative control group.The sensitivity results of duplex PCR showed that distinct target amplicons could still be visualized when viral templates were diluted up to 512-fold.In conclusion,the established duplex PCR system can simultaneously detect SeV and PVM in mixed samples,exhibiting strong specificity and high sensitivity.It can be applied to disease prevention and pathogen monitoring.关键词
仙台病毒/小鼠肺炎病毒/双重PCRKey words
Sendai virus/Pneumonia virus of mice/duplex PCR分类
农业科技引用本文复制引用
田永刚,张景科,王姗姗,马旭炅,段天林..啮齿类实验动物仙台病毒和小鼠肺炎病毒双重PCR检测方法的建立[J].中兽医医药杂志,2026,45(5):71-76,6.基金项目
甘肃省自然科学基金实验动物专项(24JRRA475) （24JRRA475）
