快速检测汉滩病毒新型实时荧光定量RT-PCR方法的建立与应用  

Establishment of quantitative real-time RT-PCR of hantaan virus

作　　者：何芳 杨鹏飞[1] 陈国清[2] 唐丽[1] 燕清丽[1] 刘纯成[1] 赵怀荣[1] HE Fang;YANG Peng-fei;CHEN Guo-qing;TANG Li;YAN Qing-li;LIU Chun-cheng;ZHAO Huai-rong(Huai'an Center for Disease Control and Prevention, Huai'an 223001, Jiangsu, China)

机构地区：[1]淮安市疾病预防控制中心,江苏淮安223001 [2]盐城市疾病预防控制中心

出　　处：《中国校医》2019年第6期404-406,共3页Chinese Journal of School Doctor

基　　金：淮安市重点研发计划(HAS201616);淮安市自然科学研究计划(HAB201736)

摘　　要：目的建立一种汉滩型汉坦病毒(Hantaan virus,HTNV)荧光定量(RT-PCR)的检测方法,以便快速准确地检测HTNV。方法利用Beacon Designer7.0设计引物和探针,以HTNV的S基因片段为模板,进行实时荧光定量RT-PCR,评价此方法的特异性及灵敏度。结果建立的RT-PCR方法对HTNV的最低检出限为4.08 copies/μL,模板Ct值与稀释浓度的对数之间具有良好的线性关系,标准曲线方程为Y=-3.4118X+41.997,扩增效率为96.4%,R^2=0.994。结论所建立的实时荧光定量RT-PCR检测方法灵敏度高、特异性好,可应用于HTNV的快速检测。Objective To conduct a universal real-time fluorescence RT-PCR method as a rapid and accurate approach for detecting hantaan virus.Methods The twin primer and the TaqMan probe were designed and synthesized based on the S gene segment of hantaan virus.The real-time RT-PCR was developed for testing the sensitivity and specialty.Results The Ct value of templates had a linear relationship with the log starting quantity.The minimum detection limit of HTNV for RT-PCR method was 4.08 copies/μL,and the standard curve Y=-3.4118 X+41.997,the amplification efficiency was 96.4%,and R^2=0.994.Conclusion The quantitative real-time fluorescence for hantaan virus detection is established and characterized by rapidity and sensitivity.

关 键 词：汉滩型汉坦病毒 S基因片段 定量 实时荧光RT-PCR 

分 类 号：R373.9[医药卫生—病原生物学]