口蹄疫病毒RT-PCR检测与血清型鉴别

针对编码非结构蛋白的3D基因合成一对引物进行口蹄疫病毒RT-PCR扩增,不同血清型病毒的RNA样本均显现一条457bp的目的带,与预期设计的长度相符合。在敏感性试验中,O型、A型和AsiaⅠ型病毒的最小RNA检出量分别为0.8ng、8ng和8ng。根据GenBank发表的口蹄疫病毒VP1和2A基因序列,采用多重RT-PCR鉴别口蹄疫病毒血清型,O型、A型和AsiaⅠ型病毒的特异性扩增片段分别为200bp、340bp和500bp。对9份乳鼠感染病料进行检测,确诊为O血清型口蹄疫病毒感染。

The RT-PCR Detection and Serotype Differentiation of Foot-and-Mouth Disease Viruses

A pair of primers targeting the 3D gene encoding the non-structural protein was synthesized for RT-PCR amplification of foot-and-mouth disease virus (FMDV), which was located on. The RNA samples of different serotype viruses manifested 457bp target band in accordance with the expected design length. The sensitivity test showed that the minimum RNA contents of 0.8ng、8ng and 8ng could be detected for O、A and Asia Ⅰtype viruses. Based on the published nucleotide sequences of FMDV VP1 and 2A genes, multiplex RT-PCR were conducted for differentiation of FMDV serotypes. The specific amplified bands were 200bp、340bp and 500bp respectively for O、A and AsiaⅠ type viruses.9 affected baby mouse tissue samples were also detected and diagnosed as infection with FMDV O serotype.