兔出血症病毒RT-PCR检测方法的建立及应用

摘要:按照GenBank上公布的兔出血症病毒（RHDV）基因序列设计一对引物，以RHDV发病兔的新鲜肝为材料，提取总RNA，进行cDNA的合成和PCR扩增，结果能扩增出与预期的269bp大小一致的片段，该产物序列与公布的RHDV基因序列有95.8%～98.5%的同源性，表明已成功建立了RHDV RT-PCR检测方法。用该方法和血凝实验（HA）对兔出血症发病兔各组织脏器进行检测，结果表明，发病死亡兔各脏器中，HA检测阳性的病料为肝脏、肾脏、脾脏、血液、肺；RT-PCR检测除粪便外，其它均为阳性，其敏感性是HA的4×104倍。RT-PCR方法检测RHDV敏感度高、特异性强、重复性好，不仅可用于RHD临床诊断和流行病学调查，而且在兔肉等兔类产品的检疫方面具有很好的应用前景。

Establishment and Application of RT-PCR Test for Rabbit hemorrhagic disease virus

Abstract: the RT-PCR method was established. In this test, total RNA was extracted from the fresh liver of rabbit haemorrhagic disease rabbits, a pair of primers were designed according to the sequences of RHDV published in GenBank, which can amplified a 269 bp segment. The product was sequenced and there was 95.8％～98.5％homology between the product and the relevant sequence published in GenBank. RHDV viscera distribution experiment shows that liver, kidney, spleen, blood, lung were HA positive, and all the viscera except feces were positive by RT-PCR. The sensitivity of RT-PCR for RHDV is 4×104times that of HA. All those experiments above showed the established RT-PCR method detecting RHDV has strong speciality, high sensitivity and good repetition. This RT-PCR can not only be used in RHDV clinical diagnosis and epidemiology study but also in quarantine of rabbit product ,example for meat.