基于DPO引物的多重RT-PCR技术检测常见呼吸道病毒  

Detection of common respiratory viruses by multiplex RT-PCR and DPO primers

作　　者：徐昌平[1] 卢亦愚[1] 冯燕[1] 陈寅[1] 钟淑玲[1] 

机构地区：[1]浙江省疾病预防控制中心,杭州310051

出　　处：《中国卫生检验杂志》2013年第3期658-660,共3页Chinese Journal of Health Laboratory Technology

基　　金：浙江省医药卫生科学研究基金(2008A027);浙江省医学重点学科群(XKQ-009-003);国家科技重大专项-浙江及周边省传染病病原谱流行规律研究(2012ZX10004-210-002)

摘　　要：目的:应用DPO引物技术建立一种同时检测流感等5种常见呼吸道病毒的多重RT-PCR检测方法。方法:设计针对流感病毒甲、乙型,呼吸道合胞病毒,腺病毒,以及肠道病毒通用的高特异性DPO引物,通过对多重RT-PCR反应体系的优化,建立上述5种病原体的多重RT-PCR检测技术。结果:建立的多重RT-PCR方法可同时扩增甲型流感病毒等5种病原体的基因片段,检测敏感性分别为102copies/ml、103copies/ml、102copies/ml、103copies/ml、102copies/ml,并与副流感等其它呼吸道病毒无交叉反应。结论:本研究建立的多重RT-PCR可以同时准确分型甲、乙型流感病毒,呼吸道合胞病毒,腺病毒和肠道病毒,为呼吸道感染的病原学诊断提供了有效的实验室检测手段。Objective： To establish a multiplex RT - PCR assay for detection of five important respiratory viruses using the dual priming oligonucleotide system. Methods ： Five sets of dual priming oligonucleotide primers for influenza A virus, influenza B virus, respiratory syncytial virus, adenovirus, enteroviruses were designed, and a multiplex RT - PCR technique was developed to simultaneously detect these five viruses. Results： Five specific PCR products were amplified. The specificity of the technique was verified, and the sensitivity for these five pathogens were 10^2 copies/ml, 10^3 copies/ml, 10^2 copies/ml, 10^3 copies/ml, 10^2 copies/ml respectively. Conclusion： This assay was a rapid, sensitive and specific method for simultaneous detection of influenza A virus, influenza B virus, respiratory syncytial virus, adenovirus and enteroviruses.

关 键 词：呼吸道病毒 DPO引物 多重RT—PCR 

分 类 号：R446.5[医药卫生—诊断学]