PEDV疫苗株与野毒株RT-PCR鉴别诊断方法的建立及临床应用

Development and Clinical Application of RT-PCR Differential Diagnosis Method for Vaccine Strains and Wild Strains of PEDV

根据猪流行性腹泻病毒(PEDV)疫苗株和野毒株在ORF3基因上的差异,我们设计了一对引物用于建立PEDV疫苗株和野毒株的RT-PCR鉴别诊断方法,其中疫苗株PCR扩增产物长度为213 bp,野毒株为262 bp。特异性试验结果表明,该引物对猪传染性胃肠炎病毒、猪轮状病毒、猪瘟病毒、猪伪狂犬病病毒和猪繁殖与呼吸综合征病毒的扩增均为阴性。2014年3月至2015年4月期间利用该方法对河南省51家猪场的581份腹泻样本进行检测,结果发现,PEDV总阳性率为54.9%(391/581),其中野毒株阳性率84.9%(332/391),疫苗株阳性率为15.1%(59/391),说明当前腹泻样本中的PEDV检出率依然较高,且在这些PEDV阳性病料中以野毒感染为主。

Based on the genetic differences of ORF3 of the pig epidemic diarrhea virus（PEDV） vaccine strains and wild strains,we designed a pair of primers to establish a RT-PCR method for the differential diagnosis of PEDV vaccine strains and wild strains,of which the PCR amplification product length of the vaccine strains was 213 bp,and the wild strains was 262 bp.Specific test results showed that the amplification reactions of the primers to the pig infectious gastroenteritis virus,porcine rotavirus,swine fever virus,pig pseudo rabies virus and porcine reproductive and respiratory syndrome virus were negative.We used this method to test 581 diarrhea samples from 51 pig farms of Henan province during the period of March 2014 to April 2015,and the results showed that the total positive rate of PEDV was 54.9%（391/581）,of which the positive rate of wild strains was 84.9%（332/391）,and the vaccine strains was 15.1%（59/391）.All of these results indicate that the detection rate of PEDV in the current diarrhea sample is still high,and the PEDV positive samples mainly are wild strain infections.