猪繁殖与呼吸综合征病毒变异株和经典株RT-PCR鉴别诊断技术及其临床应用

【目的】建立能针对猪繁殖与呼吸综合征病毒（PRRSV）经典株和变异株的RT-PCR鉴别诊断技术,为有效防控猪繁殖与呼吸综合征（PRRS）提供技术支撑。【方法】根据GenBank中已发表的PRRSV Nsp2基因序列,设计1对特异性引物,建立能鉴别PRRSV经典株和变异株的RT-PCR诊断技术,并进行特异性、敏感性试验及临床应用。【结果】建立的RT-PCR能同时扩增获得两条大小与预期结果一致的特异片段,即332bp（经典株）和241bp（变异株）,根据其大小可将两者鉴别区分;而且具有较强的特异性和较高的敏感性,能同时检测出100fg PRRSV经典株和变异株的RNA含量;临床应用发现37份可疑病料中PRRSV经典株占24.32%,变异株占51.35%,其余样品为阴性。【结论】该RT-PCR技术能有效鉴别PRRSV经典株和变异株,且具有快速简便、特异敏感的特点,在临床诊断方面具有广阔的应用前景。

RT-PCR differential diagnosis technique for variant and classical strains of PRRSV and its clinical application

Objective]The studies were conducted to establish the RT-PCR assay for the differential diagnosis of the highly pathogenic porcine reproductive and respiratory syndrome virus (PRRSV)(variant strain) and classical PRRSV in order to provide control measures for PRRS. Method]A pairs of specific primers was designed on the basis of Nsp2 gene sequences of PRRSV collected from GenBank to amplify PRRSV genes using RT-PCR for differentiating the variant and classical strains of PRRSV. The specificity and sensitiv...