[从人类临床标本中检测H5N1的方法的开发]
[Development of methods for detection of H5N1 from human clinical specimens].
作者信息
Wen Le-ying, Xu Hong, Lan Yu, Zhao Xiang, Zhang Xiao-guang, Wang Da-yan, Yao Li-hong, Dong Jie, Zhang Jia-huai, Guo Yuan-ji, Shu Yue-long
机构信息
Chinese Influenza Center, Institute for Viral Disease Control and Prevention, State Key Laboratory for Infectious Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 100052, China.
出版信息
Zhonghua Shi Yan He Lin Chuang Bing Du Xue Za Zhi. 2006 Jun;20(2):24-6.
BACKGROUND
To establish a method for H5N1 RNA detection and laboratory diagnosis of suspected human avian influenza (H5N1) virus infected cases.
METHODS
The typing and sub-typing primers were designed according to M and H5 and N1 gene respectively, and the RT-PCR and real-time PCR were developed using these primers.
RESULTS
The RT-PCR and real-time PCR could be used for H5N1 detection specifically, and there was no cross reaction with other influenza subtypes such as H1 and H3. The sensitivity for RT-PCR and real-time PCR was 1 TCID50 and 0.01 TCID50 respectively. Thirteen laboratory confirmed human H5N1 cases were detected from 42 suspected cases by using these methods.
CONCLUSION
The established RT-PCR and real-time PCR methods can be used for laboratory detection of suspected human H5N1 cases.
背景
建立一种用于检测H5N1 RNA及对疑似人感染H5N1禽流感病毒病例进行实验室诊断的方法。
方法
分别根据M基因、H5基因和N1基因设计分型及亚型引物，并使用这些引物开展逆转录聚合酶链反应（RT-PCR）和实时荧光定量聚合酶链反应（real-time PCR）。
结果
RT-PCR和real-time PCR可特异性用于H5N1检测，与H1、H3等其他流感亚型无交叉反应。RT-PCR和real-time PCR的灵敏度分别为1半数组织培养感染剂量（TCID50）和0.01 TCID50。使用这些方法从42例疑似病例中检测出13例实验室确诊的人感染H5N1病例。
结论
所建立的RT-PCR和real-time PCR方法可用于疑似人感染H5N1病例的实验室检测。
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