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  

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

作　　者：梁民 蒋柳平[2] 黄溢泓[2] 盘美妮[2] 

机构地区：[1]柳州市水产畜牧兽医局 [2]柳州市动物疫病预防控制中心,广西柳州545005

出　　处：《南方农业学报》2011年第8期991-994,共4页Journal of Southern Agriculture

基　　金：广西柳州市科学基金项目(2008060605)

摘　　要：【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。[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 sensitivity test were performed and clinical applications were carried out. [Result]The fragments of 332 and 241 bp were amplified from classical and variant strains of PRRSV using RT-PCR, respectively, which could be differentiated on the basis of their sizes. The sensitivity test results demonstrated that as little as 100 fg of the classical strain of PRRSV RNA, and 10 fg of the variant strain of PRRSV RNA could be detected by RT-PCR. The clinical application of RT-PCR showed that, out of 37 suspicious disease samples, 24.32 and 51.35% belonged to classical and variant strains of PRRSV, respectively, while the others showed negative. [Conclusion]The established RT-PCR differential diagnosis technique was found to differentiate the classical and variant strains of PRRSV effectively, and showed highly specific and simple assay, which may be widely used in clinical diagnosis.

关 键 词：猪繁殖与呼吸综合征病毒(PRRSV) 经典株 变异株 RT-PCR 

分 类 号：S858.28[农业科学—临床兽医学] S854.44[农业科学—兽医学]