多重RT-PCR检测猪口蹄疫病毒、猪水泡病病毒和猪水疱性口炎病毒的研究  

Simultaneous Detection of Foot-and-mouth Disease Virus,Swine Vesicular Disease Virus and Vesicular Stomatitis Virus by Multiplex RT-PCR

作　　者：祁会彩[1] 龚振华[1] 杨增岐[2] 郑增忍[1] 潘康锁 李葳[1] 黄秀梅[1] 郭福生[1] 蒋正军[1] 

机构地区：[1]农业部动物检疫所,山东青岛266032 [2]西北农林科技大学动物科技学院,陕西杨凌712100 [3]陕西省畜牧兽医总站,西安710016

出　　处：《中国动物检疫》2006年第3期24-26,共3页China Animal Health Inspection

基　　金：人畜共患疾病及植物疫病病原体检测技术(2001BA804A22)

摘　　要：建立了一种同时检测猪口蹄疫病毒(FMDV)、猪水泡病病毒(SVDV)和猪水疱性口炎病毒(VSV)三种病原体的多重RT-PCR方法。参照文献报道的基因序列,设计合成了三对特异性引物;PCR扩增条件进行优化后,用这三对引物对同一样品中的FMDV、SVDV、VSVRNA模板进行扩增,结果同时得到了三条特异性条带,大小与试验设计相符:FMDV(208bp)、SVDV(862bp)、VSV(638bp),且对猪瘟病毒(CSFV)、猪繁殖与呼吸综合症病毒(PRRSV)和猪传染性胃肠炎病毒(TGEV)核酸扩增结果为阴性;三种病毒RNA模板检出的最小量均为10fg。试验证明,此方法经济、快速、敏感、特异,可用于FMDV、SVDV和VSV这三种猪水泡性疾病的鉴别诊断及流行病学调查。A multiplex RT-PCR was optimized to simultaneously detect foot-and-mouth disease virus （FMDV）, swine vesicular disease virus （SVDV） and vesicular stomatitis virus （VSV）. Three specific primer sets were selected according to FMDV, SVDV, and VSV gene sequences reported. The results showed that all the＇ Sampies which contained FMDV, SVDV and VSV could be amplified by this method with optimized factor of reaction. Three specific bands of FMDV 208 bp, and SVDV 862 bp, and VSV 638 bp, as low as i0fg of FMDV-RNA, SVDV-RNA, and VSV-RNA could be detected and visualized under UV light, but not the gene fragments of PRRSV,TGEV and CSFV. The multiplex PCR appears to be rapid, sensitive and Specific method for detecting FMDV, SVDV and VSV simultaneously, and may be applied to clinical diagnosis and epidemiological investigation.

关 键 词：多重PCR 猪口蹄疫病毒 猪水泡病病毒 猪水疱性口炎病毒 

分 类 号：S858.285.3[农业科学—临床兽医学] S858.291[农业科学—兽医学]