目的对一例不明原因重症肺炎病例进行H5N6禽流感病毒实验室诊断。方法采集患者呼吸道标本,利用两种核酸提取方法,使用实时荧光RT-PCR技术进行检测。结果 H5和N6亚型引物的检测结果均为阳性,其中仪器提取法扩增曲线较手工提取法扩增曲线结果典型。结论该病例经实验室检测,为湖南省湘西自治州首例H5N6禽流感病例。为提高H5N6禽流感病毒的检出率,建议在可疑病例使用药物之前,及早采集下呼吸道标本送实验室检测。 Objective To diagnose H5N6 bird flu virus in a case of unexplained severe pneumonia. Methods The respiratory specimens of patients were collected and detected by real-time fluorescence RT-PCR using two nucleic acid extraction methods. Results The detection results of H5 and N6 subtypes were all positive. The amplification curve of instrument extraction method was more typical than that of manual extraction method. Conclusion The case was detected by laboratory and was the first H5N6 bird flu case in Xiangxi Autonomous Prefecture of Hunan Province. In order to increase the detection rate of H5N6 bird flu virus, it is suggested that lower respiratory tract specimens be collected and sent to laboratory for testing before use of drugs in suspected cases.