猪繁殖与呼吸综合征病毒不同毒株RT-PCR鉴别诊断方法的建立  

Establishment of differential diagnosis methods for different strains of porcine reproductive and respiratory syndrome virus by RT-PCR

作　　者：孟令宅 张哲[1] 刘陈针玉 赵卓[2,3] 刘涛 袁万哲[1,5] MENG Ling-zhai;ZHANG Zhe;LIU CHen-zhenyu;ZHAO Zhuo;LIU Tao;YUAN Wan-zhe(College of Animal Medicine,Hebei Agricultural University,Baoding 071001,China;Beijing Ecological Technology Co.,Ltd,Beijing 102600,China;;Beijing Centrebio Biological Co.,Ltd,Beijing 462629,China;Shijiazhuang City Animal Health Super Vision Office,Shijiazhuang 050000,China;Hebei Engineering and Technology Research Center of Veterinary Biotechnology,Baoding 071001,China)

机构地区：[1]河北农业大学动物医学院,河北保定071001 [2]北京生泰尔科技股份有限公司,北京102600 [3]北京华夏兴洋生物科技有限公司,北京102629 [4]石家庄市动物卫生监督所,河北石家庄050000 [5]河北省兽医生物技术工程技术研究中心,河北保定071001

出　　处：《河北农业大学学报》2018年第6期98-103,共6页Journal of Hebei Agricultural University

基　　金：河北省高校百名优秀创新人才支持计划(Ⅲ)(SLRC2017039)

摘　　要：根据GenBank登录的高致病性猪繁殖与呼吸综合征病毒(HP-PRRSV)、经典型猪繁殖与呼吸综合征病毒(C-PRRSV)、与NADC 30-like猪繁殖与呼吸综合征病毒(NADC 30-like PRRSV)Nsp2基因的核苷酸序列,设计了1对用于扩增3种不同毒株的RT-PCR引物,建立了HP-PRRSV、C-PRRSV、NADC 30-like PRRSV等3种毒株RT-PCR鉴别诊断的方法,该方法可扩增出1072bp(C-PRRSV),985bp(HP-PRRSV)与682bp(NADC 30-like PRRSV)的特异性片段,而对猪瘟病毒,伪狂犬病毒与猪圆环病毒2型检测均为阴性。敏感性试验结果表明,对HP-PRRSV,C-PRRSV、NADC 30-like PRRSV的最低核酸检出量分别为385,269,402pg,对来自临床的39份组织样品进行检测,检测阳性率为51%。该方法可用于3种不同PRRSV毒株临床快速检测与流行病学调查。According to the Nsp2 gene sequences of Highly pathogenic porcine reproductive and respiratory syndrome virus(HP-PRRSV),classical porcine reproductive and respiratory syndrome virus(C-PRRSV),and NADC 30-like porcine reproductive and respiratory syndrome virus(NADC 30-like PRRSV)published in GenBank.A pair of RT-PCR primers was designed to amplify three different strains.A assay for detection of three strains of HP-PRRSV,C-PRRSV and NADC 30-like PRRSV by RT-PCR was established.Specific PCR products of 1072 bp(C-PRRSV),985 bp(HP-PRRSV)and 682 bp(NADC 30-like-PRRSV)could be amplified by this assay.However,the testing results for swine fever virus,pseudorabies virus and porcine circovirus type 2 were all negative.The sensitivity test results showed that the minimum nucleic acid detection levels for HP-PRRSV,C-PRRSV,and NADC 30-like PRRSV were 385,269 and 402 pg.Detecting 39 clinical samples for clinical testing,the positive rate is 51%.The assay could be used for clinical rapid detection and epidemiological investigation of three different PRRSV strains.

关 键 词：猪繁殖与呼吸综合征病毒 高致病性 经典型 NADC 30-like 鉴别 

分 类 号：S852.65[农业科学—基础兽医学]