成人重症肺炎患者EB病毒和人鼻病毒混合感染的病原学分子鉴定

目的对1例有禽类接触史的成人重症肺炎患者进行病原学研究,确定其病原体。方法采集患者的下呼吸道标本,应用荧光PCR方法检测禽流感H5N1和SARS病毒及甲型H1N1和季节性流感病毒;应用多重PCR方法检测12种常见呼吸道病毒;应用普通PCR方法检测EB病毒和巨细胞病毒,扩增阳性的病毒基因片段测序后,与GenBank数据库参考序列进行同源性分析。结果荧光PCR法检测禽流感病毒H5N1病毒和SARS病毒核酸阴性,甲型H1N1和季节性流感病毒核酸阴性;多重PCR检测人鼻病毒、EB病毒均阳性;普通PCR检测EB病毒核酸阳性。基因序列比对显示,EB病毒核苷酸序列与GenBank数据库中参考序列同源性为99.5%～100%;人鼻病毒的核苷酸序列与GenBank数据库中A组鼻病毒(73血清型)同源性最高,为94.7%。结论该例重症肺炎患者为EB病毒和A组(73血清型)人鼻病毒混合感染,排除禽流感H5N1和SARS感染。

Molecular identification of a mixed infection of EB and human rhinovirus in an adult patient with severe pneumonia

Objective To determine the etiology of severe pneumonia in an adult patient with a history of contact with birds.Methods A lower respiratory tract specimen was collected from the patient to extract nucleic acid for use as a template in real-time PCR.Multiplex PCR was used to test for H5N1,SARS-CoV,pandemic H1N1,seasonal influenza virus,and other common respiratory viruses and unusual pathogens including EB and CMV virus.Resulting gene fragments were identified and their homology was analyzed with other reference sequences downloaded from the GenBank database.Results TPCR revealed that the patient had a mixed infection of the EB virus and human rhinovirus.Nucleotide sequences had a homology as high as 99.5%-100% with the EB virus and 94.7% with human rhinovirus(A-serotype 73).Conclusion Based on laboratory analysis,the patient had a mixed infection of the EB virus and human Rhinovirus(A-serotype 73).The possibility of infection with the H5N1 virus or SARS was excluded..