中国动物传染病学报2025，Vol.33Issue(3)：69-74,6.
伪狂犬病病毒gE基因SYBR GreenⅠ实时荧光定量PCR检测方法的建立
Development of SYBR Green Ⅰ Real-Time Quantitative PCR to Detect of gE Gene of Porcine Pseudorabies Virus
摘要
Abstract
In the study,a pair of specific primers were designed by comparing the gE gene of Porcine pseudorabies virus prevalent strain with that of classical strain and a real-time quantitative PCR(qPCR)method was developed for rapid detection of Porcine pseudorabies virus.The results showed that the qPCR method had good repeatability and the coefficients of variation within and between batches were 0.62%-1.30%,0.47%-1.36%.The qPCR method had high specificity and had no amplification reaction with Japanese encephalitis virus,Porcine epidemic diarrhea virus,Porcine reproductive and respiratory syndrome virus,Porcine delta coronavirus,Porcine parvovirus and Porcine pseudorabies vaccine strains.The sensitivity test showed that the lowest template copy concentration was 100 copies/μL by the qPCR method,which was 100 times higher than that of conventional PCR.Subsequently,76 clinical samples were tested with both qPCR and conventional PCR and positive rates were 42.11%(32/76)by the qPCR method and 36.84%(28/76)by conventional PCR,indicating the coincidence rate at 95%.Furthermore,4 PCR amplification products that were positive by qPCR and negative by conventional PCR were sequenced and confirmed to be gE target gene fragments.Therefore,the qPCR method developed in this study was a rapid and accurate tool for the detection and differentiation of field and vaccine strains of Porcine pseudorabies virus.关键词
伪狂犬病病毒/gE基因/SYBR GreenⅠ/荧光定量PCRKey words
Porcine pseudorabies virus/gE gene/SYBR Green I/real-time quantitative PCR分类
农业科技引用本文复制引用
邓鑫,伍锐,魏博文,彭文婧,颜其贵,赵勤,黄小波,文翼平,杜森焱,曹三杰..伪狂犬病病毒gE基因SYBR GreenⅠ实时荧光定量PCR检测方法的建立[J].中国动物传染病学报,2025,33(3):69-74,6.基金项目
国家重点研发项目(2023YFF0724501) （2023YFF0724501）
