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

发布时间：2018-02-25 17:43

本文关键词： 猪繁殖与呼吸综合征病毒美洲型多重RT-PCR 野毒株疫苗株　出处：《中国畜牧兽医》2017年03期 　论文类型：期刊论文

【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。
[Abstract]:In order to establish a rapid identification and detection method for porcine reproductive and respiratory syndrome virus (PRRSVV) of porcine reproductive and respiratory syndrome virus, this study aimed at the Nsp2 gene sequence characteristics of PRRSV American classic strain, highly pathogenic variant strain and TJM-F92 vaccine strain. Two pairs of specific primers were designed. After optimizing the reaction conditions, a multiplex RT-PCR method was established to simultaneously detect and distinguish the classical PRRSV strains, variant strains and vaccine strains. The method was highly specific and had no cross reaction with other porcine viruses. The detection limit of the standard plasmid was 1.13 脳 10 ~ 3 copies / 渭 L. the results showed that there were 119 PRRSV positive samples, 5 of which were American classical strains, using the established method to detect 349 clinical suspected samples, and the results were as follows: (1) the detection limit of the recombinant plasmid was 1.13 脳 10 ~ 3 copies / 渭 L. There were 7 strains of TJM-F92 vaccine strains and 7 strains of mixed positive strains of PRRSV and TJM-F92 vaccine strains. The results showed that the method of identification and detection of PRRSV American classic strains, variant strains and TJM-F92 vaccine strains was successfully established in this study. It can be used for clinical detection and epidemiological investigation of PRRSV.
【作者单位】：广西动物疫病预防控制中心;广西大学动物科学技术学院;
【基金】：广西科学研究与技术开发计划项目(桂科转14125004-22)
【分类号】：S852.65

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