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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。
Objective To establish a rapid and accurate method for the rapid and accurate detection of 2013 new H7N9 avian influenza virus in throat swab samples from febrile patients. Methods The universal primers were designed for the M gene of influenza A virus. Specific primers were designed according to the HA and NA genes of the 2013 H7N9 avian influenza virus. The primers were designed by using beta-actin as a human housekeeping gene. Method quadruple RT-PCR reaction system. Methods and commercial real-time fluorescence RT-PCR kit for comparison. Results The detection of 2013 new H7N9 bird flu virus was successfully established by four-step one-step RT-PCR. Conclusion The method is simple, practical and low cost and suitable for the rapid detection of 2013 new H7N9 bird flu virus.