莫秋华1，罗宝正2，杜田3，赵俊华1，祝琰2，王琪1，滕勇勇4，林继灿1，杨泽1

    1.珠海国际旅行卫生保健中心，广东珠海519020；2.珠海出入境检验检疫局；

    3.深圳市福田区疾病预防控制中心；4.南方医科大学

   摘要：目的建立能快速准确检测发热病人咽拭子样本中2013新型H7N9禽流感病毒的四重RT-PCR方法。方法针对甲型流感病毒的M基因设计通用引物，针对2013新型H7N9禽流感病毒的HA和NA基因设计特异性引物，选择人源看家基因beta-actin设计质控引物，通过优化实验条件建立一步法四重RT-PCR反应体系。与商品化实时荧光RT-PCR试剂盒进行方法比对。结果成功建立四重一步法RT-PCR筛查2013新型H7N9禽流感病毒的检测技术。结论该方法简便、实用、成本低廉，适用于2013新型H7N9禽流感病毒的快速检测。

    关键词：多重RT-PCR；甲型(H7N9)禽流感病毒；甲型流感病毒；

    中图分类号：R511.7　文献标识码：B

    Four multiplex RT-PCR for rapid detection of 2013novel

    avian influenza A (H7N9) virus

    MO Qiu-hua*, LUO Bao-zheng, DU Tian, ZHAO Jun-hua,ZHU Yan, WANG Qi, TENG Yong-yong, LIN Ji-can, YANG Ze

    *Zhuhai International Travel Health Care Center,Zhuhai， Guangdong 519020, China

    Abstract:   Objective   To establish a fourmultiplex RT-PCR assay for rapid and accurate detection of 2013novel avian influenza A (H7N9) virus in throat-swab specimens ofpatients with fever.   Methods   The universal primersfor amplifying the M gene were designed for detection of influenzaA viruses. Two pairs of specific primers of HA and NA gene weredesigned to detect the 2013 novel A (H7N9) influenza virus. Theanthropogenic housekeeping gene of beta-actin was selected andprimers were designed as quality control for throat-swab samples.The reaction system of one-step four multiplex RT-PCR assay wasestablished by optimizing the experimental conditions. Thecommercially available real-time fluorescent RT-PCR kits were usedto validate this method.   Results   The one-step fourmultiplex RT-PCR assay for screening of 2013 novel avian influenzaA (H7N9) virus was developed successfully.   Conclusion  The new method is simple, practical, low-cost and very suitable forrapid detection of 2013 novel avian influenza A (H7N9) virus.

    Key words:   Multiplex RT-PCR; Avian influenza A(H7N9) virus; Influenza A virus

    《中国国境卫生检疫杂志》2013年10月刊