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目的建立快速检测甲型H3N2流感病毒神经氨酸酶(NA)奥司他韦耐药突变位点的实时RT-PCR方法。方法通过设计特异性靶向NAR292K突变位点的Taq-MGB探针进行实时RT-PCR反应,并利用模拟样本和临床标本进行灵敏性和特异性评价。结果与结论建立了快速检测NAR292K位点的实时RT-PCR检测方法;本方法灵敏性高,可检测低至500拷贝/μl;特异性好,与无耐药突变的甲型H3N2流感病毒及其他呼吸道病毒均无交叉反应,为流感耐药监测提供了有效工具。
Objective To establish a real-time RT-PCR method for the rapid detection of oseltamivir resistance of neuraminidase (NA) of influenza A (H3N2) virus. Methods Real-time RT-PCR was performed by designing a Taq-MGB probe that specifically targets the NAR292K mutation site. Sensitivity and specificity were assessed using simulated samples and clinical specimens. RESULTS AND CONCLUSION: A real-time RT-PCR method for rapid detection of NAR292K locus was established. The method is sensitive and can detect as little as 500 copies / μl. The method is characterized by good specificity and resistance to the H3N2 influenza A virus and other No cross-reaction of respiratory viruses, influenza drug resistance monitoring provides an effective tool.