多重荧光RT-PCR技术检测猪流感病毒、猪乙型脑炎病毒、猪瘟病毒、猪繁殖与呼吸综合征病毒方法的建立  

Development of a Multiplex Fluorescent RT-PCR for Detection of 4 Swine Viruses

作　　者：梁晓艳 万东山[1,2] 郭兰英 

机构地区：[1]华美生物工程有限公司,河南洛阳471003 [2]郑州大学,河南郑州450001

出　　处：《中国动物检疫》2015年第4期68-71,共4页China Animal Health Inspection

基　　金：洛阳市科技发展计划项目;编号:1301068A

摘　　要：[目的 ]建立能同时鉴测猪流感病毒(SIV)、猪乙型脑炎病毒(JEV)、猪瘟病毒(CSFV)、猪繁殖与呼吸综合征病毒(PRRSV)的多重荧光RT-PCR方法。[方法 ]根据4种病原体的高度保守基因序列,设计相应引物和探针,并对引物和探针浓度进行优化,检测48例疑似样本。[结果 ]建立了多重荧光RT-PCR方法,检测48例临床样本中,4种病毒均未感染的1例,SIV与JEV混合感染占2.1%,CSFV与PRRSV混合感染为31.3%。[结论 ]多重荧光RT-PCR对四种病原体的检测准确性好,具有快速的特点。[Objective] To develop a multiplex fluorescent RT-PCR assay to detect swine influenza virus（SIV）,Japanese encephalitis virus（JEV）,classical swine fever virus（CSFV）,and porcine reproductive and respiratory syndrome virus（PRRSV）. [Method] Specific primers and probe were designed based on the highly conserved gene sequences of the four swine pathogens and their working concentrations and the amplification conditions were optimized. The multiplex fluorescent RT-PCR assay was evaluated with 48 suspected clinical samples. [Result] The multiple fluorescent RT-PCR assay was established for 4 swine viruses. Only one out of 48 samples was negative to the four viruses;the positive rate of SIV and JEV mixed infection was 2.1 %;the positive rate of CSFV and PRRSV mixed infection was 31.3 %.[Conclusion] The development of the multiplex fluorescent RT-PCR provided a sensitive,specific and rapid method to detect SIV,CSFV,PRRSV and JEV.

关 键 词：多重荧光RT-PCR 猪流感病毒 猪乙型脑炎病毒 猪瘟病毒 猪繁殖与呼吸综合征病毒 

分 类 号：S854.44[农业科学—临床兽医学]