基于弓形虫529-bp重复序列巢式PCR诊断方法的建立

Establishment of nested PCR based on 529-bp repetitive sequence for diagnosis of Toxoplasma gondii

根据GenBank已发表的弓形虫529-bp重复序列(AF146527)设计巢式PCR引物,建立巢式PCR检测方法。结果表明,该方法能扩增出427bp的片段,敏感性试验表明该方法可以检测出0.1pg的弓形虫基因组DNA,敏感性是常规PCR的100倍。巢式PCR方法对蜥蜴利什曼原虫、隐孢子虫、细粒棘球绦虫基因组扩增无条带,特异性强。样品检测符合率达100%。巢式PCR方法的建立为弓形虫病的诊断及流行病学调查提供技术支持。

The nested PCR primers were designed according to GenBank published 529-bp repetitive DNA sequence（AF146527） in To：coplasma gondii ,we have established the nested PCR. The prod- uct of the nested PCR was about 427 bp and its sensitivity was 100 times than that of conventional PCR,which could detect the genomic DNA as low as 0.1 pg. Detection of Leishmania tarentolae, Cryptosporidium parvum and Echinococcus granulosus genome DNA by nested PCR had no fragment. The diagnosis coincidence rate is 100%. The nested PCR method is dependable for diagnosis of Toacoplasma gondii.