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首页> 外文期刊>Journal of Virological Methods >A rapid RT-PCR method to differentiate six established genotypes of rabies and rabies-related viruses using TaqMan technology.
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A rapid RT-PCR method to differentiate six established genotypes of rabies and rabies-related viruses using TaqMan technology.

机译：一种快速的RT-PCR方法，使用TaqMan技术区分狂犬病和狂犬病相关病毒的六种已确定基因型。

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摘要

A rapid and sensitive reverse transcriptase-polymerase chain reaction (RT-PCR) assay incorporating TaqMan probes has been developed that can distinguish among the six established rabies and rabies-related virus genotypes. TaqMan probes were designed and validated against 106 rabies and rabies-related virus isolates, one isolate of the Australian bat Lyssaviruses (genotype 7), and 18 other non-rabies viruses important in the veterinary field. The N gene was used as the target for the probes as it is well conserved and has been intensively used to genotype rabies isolates. Additionally, it was found to contain regions specific to each genotype conducive to probe design. The RT-PCR assay described amplifies a portion of the nucleoprotein gene of all 107 rabies and rabies-related viruses, but none of the other viruses tested. Inclusion of TaqMan-genotype-specific probes in the RT-PCR assay permits rapid identification of the virus present. By combining RT-PCR with TaqMan genotyping probes suspect rabies virus isolates can be identified in a single closed tube system that prevents potential PCR-product carry over contamination.
机译：已经开发出一种结合TaqMan探针的快速灵敏的逆转录酶-聚合酶链反应（RT-PCR）分析方法，该方法可以区分六种已建立的狂犬病和与狂犬病相关的病毒基因型。 TaqMan探针针对106种狂犬病和狂犬病相关病毒分离株，一种澳大利亚蝙蝠狂犬病病毒（基因型7）分离株以及18种在兽医领域重要的非狂犬病病毒进行了设计和验证。由于N基因保守性好，已被广泛用作狂犬病毒分离株的基因型，因此它被用作探针的靶标。另外，发现其包含对每种基因型具有特异性的区域，这些区域有利于探针设计。所述的RT-PCR测定法扩增了所有107种狂犬病和狂犬病相关病毒的一部分核蛋白基因，但没有检测到其他病毒。在RT-PCR分析中加入TaqMan基因型特异性探针可以快速鉴定存在的病毒。通过将RT-PCR与TaqMan基因分型探针结合使用，可以在单个封闭管系统中鉴定出可疑狂犬病毒分离株，从而避免了潜在的PCR产物残留污染。

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