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〔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 explore suitable real-time fluorescent RT-PCR reagents for the detection of suspected influenza A (H1N1) virus at the border crossings by combining the detection primers and probes published by WHO with the H1N1 influenza virus. [Methods] The sensitivity, signal intensity, reaction efficiency and other factors of the three commonly used real-time fluorescent RT-PCR reagents AB AgPath-ID, Qiagen QuantiTect and Takara One Step PrimeScript were analyzed by detecting the continuous gradient dilution positive control. [Results] The detection limits of the three reagents were all 1: 1000 dilution positive control; the linear slopes of AB AgPath-ID, Qiagen QuantiTect and Takara One Step PrimeScript were -5.517, 5.214 and -4.600, respectively; QuantiTect relative to the other two obvious weakness. [Conclusion] It provides a powerful technical guarantee for the detection and monitoring of imported H1N1 flu cases at border crossings.