畜牧与饲料科学 ›› 2015, Vol. 36 ›› Issue (2): 32-32.doi: 10.12160/j.issn.1672-5190.2015.02.012

• 基础科学 • 上一篇    下一篇

猪繁殖与呼吸综合征病毒经典株、高致病性株与疫苗株TJM-F92RT-PCR鉴别方法的建立

王宇菲[1,2] 陈超阳[2] 王鑫[2] 夏铭崎[2] 武华[2]   

  1. [1]沈阳化工大学制药与生物工程学院,辽宁沈阳110142 [2]华威特(北京)生物科技有限公司,北京100085
  • 出版日期:2015-02-20 发布日期:2015-02-20
  • 通讯作者: 王宇菲
  • 作者简介:王宇菲(1989-),女,硕士研究生,主要研究方向为动物疫苗研发及分子免疫学。 通讯作者:武华(1960-),男,研究员,博士,博士生导师,主要研究方向为动物疫苗研发及分子免疫学。

Establishment of RT-PCR Method for Discrimination of Porcine Reproductive and Respiratory Syndrome Virus Classic Strains, Highly Pathogenic Strains and Vaccine Strain TJM-F92

WANG Yu-fei, CHEN Chao-yang, WANG Xin, XIA Ming-qi, WU Hua (1.College of Pharmaceutical and Biological Engineering,Shenyang University of Chemical Technology,Shenyang 110142, China;2.Sinovet(Beijing)Biotechnology Co., Ltd., Beijing 100085,China)   

  • Online:2015-02-20 Published:2015-02-20

摘要: 根据GenBank公布的猪繁殖与呼吸综合征病毒经典株(LP-PRRSV CH-1a)、高致病性株(HPPRRSV TJ)及疫苗株(HP-PRRSV TJM-F92)的Nsp2基因核苷酸序列,设计2对特异性引物1st和2nd,建立鉴别LP-PRRSV、HP-PRRSV与HP-PRRSV TJM-F92的RT-PCR检测方法。用引物1st对LPPRRSV、HP-PRRSV与HP-PRRSV TJM-F92进行RT-PCR扩增,分别扩增587、497和497 bp的特异性片段;用引物2nd对HP-PRRSV与HP-PRRSV TJM-F92进行RT-PCR扩增,分别扩增1 004和644 bp的特异性片段,结合两步RT-PCR扩增结果可达到区分3者的目的;对RT-PCR的特异性、敏感性、重复性进行评价。结果表明,建立的RT-PCR检测方法具有特异性强、敏感性高、重复性好的特点,可用于LP-PRRSV、HP-PRRSV与HP-PRRSV TJM-F92的鉴别诊断。

Abstract: In order to establish a RT-PCR method which was able to discriminate the classic strains, highly pathogenic strains and vaccine strain TJM-F92 of porcine reproductive and respiratory syndrome virus (PRRSV), two sets of primers (1st and 2nd) were designed based on the published nucleotide sequences of Nsp2 gene of PRRSV in GenBank. The size of expected amplified fragments of LP-PRRSV, HP-PRRSV and HP-PRRSV TJM-F92 in RT-PCR assay using the 1st primers were 587,497 and 497 bp, respectively. The size of expected amplified fragments of HP-PRRSV and HP-PRRSV TJM-F92 in RT-PCR assay using the 2nd primers were 1004 and 644 bp, respectively.The discriminative diagnosis could be made according to the results of the two RT-PCR assays. Furthermore, the specificity, sensitivity and repeatability of the method were evaluated. The results showed that the established RT-PCR method had good specificity, sensitivity and repeatability and could be used to identify the classic strains, highly pathogenic strains and vaccine strain TJM-F92 of PRRSV.

中图分类号: