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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 establish a CODEHOP primer RT-PCR method for rapid detection of Hantavirus. Methods According to the amino acid sequence of L genome of different Hantavirus strains published in GenBank, a pair of primers was designed and synthesized by CODEHOP method. After the reaction conditions were optimized, a rapid detection method for all viruses in Hantavirus group was established. Sensitivity and specificity. Results The results of specific test showed that this method can specifically amplify Hantavirus and the size and sequence of the target fragment are consistent with the expected results. The detection limit of Hantavirus nucleic acid is 10 pg. Conclusion The established CODEHOP RT-PCR method is highly specific and sensitive and can be used for the detection of Hantavirus.