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。
Objective To establish a four muhiplex RT-PCR assay for rapid and accurate detection of 2013 novel avian influenza A （H7N9） virus in throat-swab specimens of patients with fever. Methods The universal primers for amplifying the M gene were designed for detection of influenza A viruses. Two pairs of specific primers of HA and NA gene were designed to detect the 2013 novel A （H7N9） influenza virus. The anthropogenic housekeeping gene of beta-actin was selected and primers were designed as quality control for throat-swab samples. The reaction system of one-step four multiplex RT-PCR assay was established by optimizing the experimental conditions. The commercially available real-time fluorescent RT-PCR kits were used to validate this method. Results The one-step four multiplex RT-PCR assay for screening of 2013 novel avian influenza A （H7N9） virus was developed successfully. Conclusion The new method is simple, practical, low-cost and very suitable for rapid detection of 2013 novel avian influenza A （H7N9） virus.
出处
《中国国境卫生检疫杂志》
CAS
2013年第5期296-299,共4页
Chinese Journal of Frontier Health and Quarantine
基金
珠海检验检疫局科技计划项目(ZH2013-12)
