荧光定量RT-PCR技术在副流感病毒检测中的应用研究

The application of realtime RT-PCR in Parainfluenza virus detection

目的:建立人副流感病毒的荧光定量RT-PCR快速检测方法。方法:根据GeneBank数据库中已有的人副流感毒株的HN基因核苷酸序列,用DNASTAR生物软件进行同源性分析比较,选出高度保守且特异的核苷酸区域,用Primer express5.0软件设计人副流感病毒的引物和TaqMan核苷酸探针。建立PCR反应体系,并优化反应条件。结果:建立了具有高度灵敏度和特异度的人副流感病毒的荧光定量PCR快速检测方法。结论:本研究建立实时荧光定量PCR检测方法可以快速的检测人副流感病毒,可以用于临床的早期诊断。

Objective：To develop fluorescence quantitative RT-PCR for rapid detection of human Parainfluenza virus.Methods： Highly conservative and specific nucleotide domain of HN gene was choosed according to the existed HN gene sequences of human Parainfluenza virus in genebank and the homology analysis of the sequences by DNASTAR software.Primers and TaqMan probe for human Parainfluenza virus were designed by Primer express 5.0 software.Experimental condition for PCR was explored and optimized.Results： A highly sensitive and specific method for rapid detection of human Parainfluenza virus by fluorescence quantitative RT-PCR was established.Conclusion： A method to rapidly detect human Parainfluenza virus by fluorescence quantitative RT-PCR has successfully developed in this study which can be used for early clinic diagnosis.