汉坦病毒群CODEHOP RT-PCR检测方法的建立

Development of CODEHOP RT-PCR to detect Hantavirus

目的建立一种能对汉坦病毒群进行快速检测的CODEHOP引物RT-PCR方法。方法根据GenBank发表的不同汉坦病毒L基因组氨基酸序列,利用CODEHOP方法设计合成一对引物,经反应条件优化,建立快速检测汉坦病毒群所有病毒的方法,并通过对标准毒株的检测评价方法的灵敏度和特异性。结果特异性试验结果显示,该方法可对汉坦病毒进行特异性扩增,目的片段大小和序列与预期结果相符,对汉坦病毒核酸的最小检出量为10pg。结论建立的CODEHOP RT-PCR方法的特异性强、灵敏度高,可用于汉坦病毒群的检测。

Objective To develop a rapid method of detecting Hantavirus by RT-PCR using consensus degenerate hybrid oligonueleotide primers （CODEHOPs）. Methods In accordance with the amino acid sequence of the L genome of differ ent hantaviruses published in GenBank, a pair of primers was designed using CODEHOP. A rapid method of detecting all kinds of hantaviruses was developed by optimizing the reaction conditions, and the specificity and sensitivity of this tech nique was evaluated using standard virus strains. Results Specificity testing indicated that this technique amplified ban taviruses, and the size of the target fragments and the sequences coincided with the anticipated results. The minimum detection of hantavirus RNA was 10 pg. Conclusion A highly specific and sensitive technique for CODEHOP PCR was established, and this technique can be used to detect Hantavirus.