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猪繁殖与呼吸综合征病毒变异株实时荧光定量RT-PCR检测方法的建立
引用本文:高诗敏,王敏功,邓瑞林,姚晓辉,王林川. 猪繁殖与呼吸综合征病毒变异株实时荧光定量RT-PCR检测方法的建立[J]. 中国兽医科学, 2011, 0(3)
作者姓名:高诗敏  王敏功  邓瑞林  姚晓辉  王林川
作者单位:华南农业大学;广东温氏集团;
摘    要:根据猪繁殖与呼吸综合征病毒(PRRSV)变异株和经典毒株的序列差异,分别设计了1对特异性鉴别引物和1条TaqMan探针,建立了检测PRRSV变异株的荧光定量RT-PCR方法,并对该方法进行了特异性、敏感性、重复性试验。结果表明,建立的方法可鉴别诊断PRRSV变异株和经典毒株,具有较高的特异性;可以检测相当于1TCID50的病毒模板,比常规RT-PCR方法的灵敏度高100倍;对同一样品进行4次重复试验,变异系数(CV)<10%,具有较好的重复性。应用建立的荧光定量RT-PCR和常规RT-PCR对13份PRRS疑似病例肺组织进行平行检测,结果荧光定量RT-PCR检出11份阳性,高于常规RT-PCR方法。本研究建立的荧光定量RT-PCR方法能检测高致病性PRRSV变异株,不仅特异性好、灵敏度高,而且快速简便,可用于变异PRRSV的临床诊断和流行病学调查。

关 键 词:猪繁殖与呼吸综合征病毒  变异株  实时荧光定量RT-PCR  

Development of real-time fluorescent quantitative RT-PCR assay for detection of porcine reproductive and respiratory syndrome virus variant strains
GAO Shi-min,WANG Min-gong,DENG Rui-lin,YAO Xiao-hui,WANG Lin-chuan. Development of real-time fluorescent quantitative RT-PCR assay for detection of porcine reproductive and respiratory syndrome virus variant strains[J]. Chinese Veterinary Science, 2011, 0(3)
Authors:GAO Shi-min  WANG Min-gong  DENG Rui-lin  YAO Xiao-hui  WANG Lin-chuan
Affiliation:GAO Shi-min1,WANG Min-gong1,DENG Rui-lin2,YAO Xiao-hui2,WANG Lin-chuan1,2(1.South China Agricultural University,Guangzhou 510642,China,2 Guangdong Wens,Yunfu 527439,China)
Abstract:To identify highly pathogenic porcine reproductive and respiratory syndrome virus(PRRSV) variants rapidly using real-time PCR technique,primers and TaqMan probe for specific PCR amplification were designed based on sequence differences between variant strains and classical strains.The fluorescent quantitative RT-PCR assay for detection of PRRSV variants was established.Its specificity,sensitivity and stability were examined.This assay had differentiate PRRSV variants.1 TCID50 of template RNA could be detect...
Keywords:porcine reproductive and respiratory syndrome virus  variant strains  real-time fluorescent quantitative RT-PCR  
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