三种实时荧光RT-PCR试剂在检测甲型H1N1流感病毒中的应用  

Application of three Kinds of RT-PCR Kit for Detection of Influenza A(H1N1)Viruses

作　　者：黄宗炎[1] 朱玉兰[1] 刘胜牙[1] 王佃鹏[1] 高朝贤[1] 董瑞玲[1] 黄彤文[1] 李政良[1] 胡孔新[2] 

机构地区：[1]深圳国际旅行卫生保健中心,深圳518033 [2]中国检验检疫科学研究院,北京100123

出　　处：《中国国境卫生检疫杂志》2010年第1期1-5,共5页Chinese Journal of Frontier Health and Quarantine

基　　金：国家"十一五"重大科技专项(2009ZX10602-02)

摘　　要：〔目的〕结合WHO公布的甲型H1N1流感病毒的检测引物和探针,探索合适的实时荧光RT-PCR试剂应用于国境口岸甲型H1N1流感病毒疑似标本的检测。〔方法〕通过对连续梯度稀释阳性对照品进行检测,从而分析AB AgPath-ID、Qiagen QuantiTect和Takara One Step PrimeScript 3种常用实时荧光RT-PCR试剂的灵敏度、信号强度、反应效率等因素。〔结果〕3种试剂的检测下限均为1:1000倍稀释阳性对照品;AB AgPath-ID、Qiagen QuantiTect和Takara One Step PrimeScript检测的线性斜率分别为-5.517、5.214和-4.600;信号强度上Qiagen QuantiTect相对于其他2种明显偏弱。〔结论〕为国境口岸输入性甲型H1N1流感病例的检测和监控提供了有力的技术保障。Objective To set up a fast and effective real time RT-PCR recipe to detect influenza A（H1N1）viruses emergent at frontier ports by comparing and assessing the effects of different types of real time RT-PCR kit combined with the probes and primers published by WHO.Method Comparing and valuating the sensitivity,signal strength and efficiency with other aspects in influenza A（H1N1）viruses detection using three different brands of real time RT-PCR kit named AgPath-ID,QuantiTect and One Step PrimeScript respectively by the method of continuous gradient dilution of positive influenza A（H1N1）virus RNA sample.Result The three kinds of kit had the same sensitivity at the lowest diluted positive RNA sample by 1：1000 fold;The Slope for AgPath-ID,QuantiTect and One Step PrimeScript was-5.517,-5.214 and-4.600 respectively.As for Signal Strength,QuantiTect was much poorer than the other two.Conclusion AgPath-ID,QuantiTect and One Step PrimeScript could all be used for detection of influenza A（H1N1）viruses.The Result of numerous samples detection based on One Step PrimeScript combined with three panels of primers and probes WHO recommended showed such real time RT-PCR recipe should allow us to react quickly and effectively during influenza A（H1N1）outbreaks at frontier ports.

关 键 词：实时荧光RT-PCR 甲型H1N1流感病毒 灵敏度 斜率 阈值循环 

分 类 号：R511.7[医药卫生—内科学]