作　　者： ; ; ; (邓瑞林）; (姚晓辉）;

机构地区： 华南农业大学

出　　处： 《中国兽医科学》 2011年第3期267-272,共6页

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。 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 detected.The sensitivity of this assay was 100 times higher than the conventional RT-PCR.Content could be absolutely quantified of virus.The variation coefficients were less than 10% based on 4 replications of each sample.13 lung samples were examined by the developed fluorescent quantitative RT-PCR and the conventional PCR,respectively,and 11 were detected to be infected with PRRSV by the real-time PCR,but only 9 were confirmed to be infected with PRRSV by the conventional PCR.These results showed that the developed fluorescent quantitative RT-PCR assay was rapid,specific,sensitive and simple for the detection of PRRSV variants and the identification of highly pathogenic PRRSV.

关 键 词： 猪繁殖与呼吸综合征病毒 变异株 实时荧光定量

领　　域： [农业科学] [农业科学] [农业科学]