四重RT-PCR快速检测2013新型H7N9禽流感病毒  

Four multiplex RT-PCR for rapid detection of 2013 novel avian influenza A(H7N9) virus

作　　者：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] 

机构地区：[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

出　　处：《中国国境卫生检疫杂志》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)

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.

关 键 词：多重RT—PCR 甲型(H7N9)禽流感病毒 甲型流感病毒 

分 类 号：R511.7[医药卫生—内科学]