Real-timePCR技术在呼吸道病毒检测中的应用与分析.docVIP

Real-time PCR技术在呼吸道病毒检测中的应用与分析 吴衍恒 ，刘洪波，师舞阳，谢颖，周日东，梁洪 （中山市疾病预防控制中心，广东省中山市，528400） [摘要] 目的 应用Real-time PCR技术检测呼吸道感染疾病的病毒性病原体，分析其在呼吸道病毒检测中的可行性。方法 提取标本核酸, 应用Real-time PCR技术同时检测流感病毒A型和B型（FluA、FluB），腺病毒（ADV）（BOV）（hRV）（RSV）（PIV1、PIV2、PIV3）（MPV）（OC43、229E、HKU1、NL63）14种呼吸道病毒。 结果 从420份呼吸道标本中检出阳性257份，检出率为61.2%，其中单一感染230份，单一感染检出率为54.8%，混合感染27份，混合感染检出率为6.4%。未查明病原体163份，未检出率为38.8%。结论 Real-time PCR技术可查明呼吸道感染病例的部分病毒性病原体，还需建立其他基于PCR技术的方法体系分析未知病原体。 [关键词] 呼吸道感染；病毒；病原体；Real-time PCR The application and analysis of Real-time PCR in the detection of respiratory viruses WU Yan-heng , LIU Hong-bo, SHI Wu-yang, et a1． ZhongShan Center for Disease Control and Prevention, ZhongShan, 528400, GuangDong, China [Abstract] Objective To detect the pathogen of the viral respiratory tract infection by Real-time PCR and analyze the feasibility of Real-time PCR. Methods Nucleic acids of specimens were extracted and 14 respiratory viruses including influenza A FIuA , influenza B FIuB ,adenovirus ADV , human Boca virus BOV , human rhinovirus HRV , respiratory syncytial virus RSV , parainfluenza virus type l, type 2, type 3 PIV1, PIV2, PIV3 , human metapneumovirus MPV and human coronavirus HCoV type 229E, type OC43, type HKU1, type NL63 were detected by Real-time PCR. Results Viruses were found in 257 samples in total, accounting for 61.2% 257/420 , of which 230 samples were found to be single infections 54.8% , 27 samples were mixed infections 6.4% . 163 samples could not be identified 38.8% . Conclusion Real-time PCR technique can be used to detect the viral pathogens of the part of respiratory tract infections. Therefore, we need establish other techniques based on PCR to analyze more unknown pathogens. [ Key words] Respiratory tract infection；Viruses；Pathogen；Real-time PCR 呼吸道感染是危害人类健康的重大疾病，在感染性疾病的死亡原因里排首位90％以上由病毒引起，引起呼吸道感染的病毒主要有流感病毒A型和B型（FluA、FluB），腺病毒（ADV）（BOV）（hRV）（RSV）（PIV1、PIV2、PIV3）（MPV）（OC43、229E、HKU1、NL63）方法固相免疫测定细胞直接免疫荧光，敏感性，，病原体涉及面迅速发展的分子生物学技术更敏感特异快速检测呼吸道病毒Real-time PCR