福建畜牧兽医2025，Vol.47Issue(1)：18-23,6.
猪伪狂犬病病毒gE基因荧光定量PCR检测方法的建立与应用
Establishment and application of fluorescent quantitative PCR method for detection of porcine pseudorabies virus
摘要
Abstract
In order to rapidly identify the epidemic strains of porcine pseudorabies virus,one pair of specific primers and probes were designed according to the region of 95 bp to 210 bp in the first half of the gE gene of porcine pseudorabies virus,and the reaction conditions were optimized to establish a Taqman fluorescence quantitative PCR assay that could identify the vaccine strain and the epidemic strains of porcine pseudorabies virus with gE deletion.The results showed that the lowest lower limit of detection of this method for pMD-PRV gE-positive standard plasmid was 4.4×101 copies/μL;the method was specific,with no cross-reactivity to com-mon viral diseases in pigs(swine fever virus,porcine reproductive and respiratory syndrome virus,porcine circovirus type 2,etc.),and the specificity of PRV epidemic strains and vaccine strains(Bartha-K61,HB2000,C,Ea strains)was better than that of a commercial fluorescent PCR kit;the coefficients of variation of intra-and inter-group reproducibility tests were less than 1.6%,which was good reproducibility.Applying the fluorescent quantitative PCR assay established in this study to 165 clinical samples,the positive rate was 2.42%(4/165),and 30 clinical samples with three known positives were selected and compared with the commercialized detection kit,with a 100%compliance rate.Therefore,the fluorescence quantitative PCR of PRV gE gene established in this study can be used for rapid diagnosis and epidemiological investigation of epidemic strains of porcine pseudorabies virus,and provide technical support for the control and purification of PR.关键词
猪伪狂犬病病毒/荧光定量PCR/鉴别诊断Key words
Pseudorabies virus/Real-time PCR/Differential diagnosis引用本文复制引用
黄喜荣,曹佳佳,邱飞铭,阮静学,江子墨,蒋桢,席梓恒,戴爱玲..猪伪狂犬病病毒gE基因荧光定量PCR检测方法的建立与应用[J].福建畜牧兽医,2025,47(1):18-23,6.基金项目
中央引导地方科技发展专项(2021L3028)、福建省科技重大专项项目(2019NZ09005)、龙岩市科技计划重大项目(2019LY1001)、福建省大学生创新创业训练计划项目(202311312014)资助. （2021L3028）
