猪伪狂犬病毒Real-time PCR检测方法的建立和弱毒疫苗病毒含量的检测  

Establishment of Real-time PCR Assay of Pseudorabies Virus and Viral Load Detection of PRV Attenuated Vaccine

作　　者：华涛[1,2,3,4] 唐波 黄江[1,2,3,4] 常晨 刘国阳 张雪花[1,2,3,4] 侯继波 张道华[1,2,3,4] HUA Tao;TANG Bo;HUANG Jiang;CHANG Chen;LIU Guo-yang;ZHANG Xue-hua;HOU Ji-bo;ZHANG Dao-hua(Institute of Veterinary Immunology&Engineering,Jiangsu Academy of Agricultural Sciences,Nanjing 210014,China;National Research Center of Engineering and Technology for Veterinary Biologicals,Jiangsu Academy of Agricultural Sciences,Nanjing 210014,China;Province-State Key Laboratory Breeding Base/Jiangsu Provincial Key Laboratory of Control Technology and Standard for Agro-product Safety and Quality,Nanjing 210014,China;Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses,Yangzhou 225009,China)

机构地区：[1]江苏省农业科学院动物免疫工程研究所,江苏南京210014 [2]江苏省农业科学院国家兽用生物制品工程技术研究中心,江苏南京210014 [3]省部共建国家重点实验室培育基地/江苏省食品质量安全重点实验室,江苏南京210014 [4]江苏省动物重要疫病与人兽共患病防控协同创新中心,江苏扬州225009

出　　处：《江西农业学报》2019年第9期73-78,共6页Acta Agriculturae Jiangxi

基　　金：江苏省农业自主创新项目[cx(17)3049]

摘　　要：根据伪狂犬病毒的gB基因设计引物,将含gB基因的质粒作为标准品,绘制标准曲线,建立了伪狂犬病毒PRV的荧光定量PCR检测方法。该检测方法具有良好的重复性、敏感性和特异性,不与PCV2、PPV、CSFV、JEV和PRRSV发生反应。用该方法对不同滴度病毒液的基因拷贝数进行检测,发现病毒滴度和病毒拷贝数之间存在较好的相关性。采用该方法对7种商品化PRV弱毒疫苗进行病毒含量检测,结果显示不同商品化弱毒疫苗的病毒含量存在显著差异,与说明书标注的病毒含量基本一致。In order to establish a real-time PCR method for detecting gB gene of PRV,the author designed a pair of primers and analyzed the real-time PCR method by different quantitive concentration of plasmids containing PRV gB gene.A standard curve was achieved.The results showed that this method had good repeatability,specificity and sensitivity,and did not react with PCV2,PPV,CSFV,JEV and PRRSV.The relationship of viral titer and gene copies of different PRV dilutions was analyzed by the method.The viral titer had a better correlation with viral copies.The method was used to detect the viral content of seven kinds of commercial attenuated PRV vaccines.The viral content of different commercial attenuated PRV vaccines was significantly different,which was basically consistent with the instructions.

关 键 词：猪伪狂犬病毒 GB基因 荧光定量PCR 病毒滴度 基因拷贝数 疫苗含量 

分 类 号：S852.651[农业科学—基础兽医学]