PRRSV美洲型经典株、变异株和疫苗株多重RT-PCR鉴别检测方法的建立及应用

doi:10.16431/j.cnki.1671-7236.2017.03.036

《中国畜牧兽医》 ›› 2017, Vol. 44 ›› Issue (3): 879-887.doi: 10.16431/j.cnki.1671-7236.2017.03.036

PRRSV美洲型经典株、变异株和疫苗株多重RT-PCR鉴别检测方法的建立及应用

施开创¹, 许心婷², 胡杰¹, 粟艳琼¹, 陆文俊¹, 陈汉忠²

1. 广西动物疫病预防控制中心, 南宁 530001;
2. 广西大学动物科学技术学院, 南宁 530005

收稿日期:2016-09-21 出版日期:2017-03-20 发布日期:2017-03-21
通讯作者: 施开创 E-mail:shikaichuang@126.com
作者简介:施开创(1968-),男,广西南宁人,博士,研究员,研究方向:动物疫病诊断与防控
基金资助:
广西科学研究与技术开发计划项目（桂科转14125004-22）

Establishment and Application of a Multiplex RT-PCR Assay for Differential Detection of Classical, Highly Pathogenic and Vaccine Strains of North American Genotype PRRSV

SHI Kai-chuang¹, XU Xin-ting², HU Jie¹, SU Yan-qiong¹, LU Wen-jun¹, CHEN Han-zhong²

1. Guangxi Center for Animal Disease Control and Prevention, Nanning 530001, China;
2. College of Animal Science and Technology, Guangxi University, Nanning 530005, China

Received:2016-09-21 Online:2017-03-20 Published:2017-03-21

摘要/Abstract

摘要：

为建立猪繁殖与呼吸综合征病毒（porcine reproductive and respiratory syndrome virus，PRRSV）的快速鉴别检测方法，本研究针对PRRSV美洲型经典株、高致病性变异株以及TJM-F92疫苗株的Nsp2基因序列特点，设计2对特异性引物。经优化反应条件后，建立了能同时检测并区分PRRSV美洲型经典株、变异株及疫苗株的多重RT-PCR方法。该方法特异性强，与猪其他病毒间不存在交叉反应；敏感性高，对重组质粒标准品的检出下限为1.13×10³拷贝/μL。应用所建立的方法对349份临床疑似病料进行检测，结果检出PRRSV阳性119份，其中美洲型经典株5份、变异株107份、TJM-F92疫苗株7份，且有变异株和TJM-F92疫苗株混合阳性7份。表明本研究成功建立了PRRSV美洲型经典株、变异株及TJM-F92疫苗株的多重RT-PCR鉴别检测方法，可用于PRRSV的临床检测及流行病学调查。

关键词: 猪繁殖与呼吸综合征病毒; 美洲型; 多重RT-PCR; 野毒株; 疫苗株

Abstract:

To establish a rapid method for differential detection of classical, highly pathogenic and TJM-F92 vaccine strains of North American genotype porcine reproductive and respiratory syndrome virus (PRRSV), a multiple RT-PCR assay was established. In this assay, two pairs of primers were designed according to the genomic sequences of classical, highly pathogenic and TJM-F92 vaccine strains of PRRSV. The assay could only detect PRRSV, but not detect CSFV, FMDV, PRV and PCV2. The detection limit of the method was as little as 1.13×10³ copies/μL of templates. The established assay was successfully used to detect 349 clinical samples and 119 samples were positive for PRRSV, of which 5 samples were positive for classical PRRSV (C-PRRSV), 107 samples for highly pathogenic PRRSV (HP-PRRSV) and 7 samples for TJM-F92 vaccine strain (V-PRRSV), while 7 samples were positive for HP-PRRSV and V-PRRSV. The results indicated that the established multiple RT-PCR assay could be used for differential detection and epidemiological investigation of PRRSV.

Key words: porcine reproductive and respiratory syndrome virus (PRRSV); North American genotype; multiplex RT-PCR; wild-type strain; vaccine strain

中图分类号:

S862.65⁺1

施开创, 许心婷, 胡杰, 粟艳琼, 陆文俊, 陈汉忠. PRRSV美洲型经典株、变异株和疫苗株多重RT-PCR鉴别检测方法的建立及应用[J]. 《中国畜牧兽医》, 2017, 44(3): 879-887.

SHI Kai-chuang, XU Xin-ting, HU Jie, SU Yan-qiong, LU Wen-jun, CHEN Han-zhong. Establishment and Application of a Multiplex RT-PCR Assay for Differential Detection of Classical, Highly Pathogenic and Vaccine Strains of North American Genotype PRRSV[J]. , 2017, 44(3): 879-887.

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PRRSV美洲型经典株、变异株和疫苗株多重RT-PCR鉴别检测方法的建立及应用

Establishment and Application of a Multiplex RT-PCR Assay for Differential Detection of Classical, Highly Pathogenic and Vaccine Strains of North American Genotype PRRSV

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