人偏肺病毒检测方法的比较  

Comparison of Various Methods for Determination of Human Metapneumovirus

作　　者：任妍[1] 陈昕[1] 杜丽娜[1] 赵晓东[1] 

机构地区：[1]重庆医科大学附属儿童医院P2实验室,重庆400014

出　　处：《中国生物制品学杂志》2010年第2期187-191,共5页Chinese Journal of Biologicals

基　　金：重庆市科技计划攻关项目(CSTC;2005AC5062);全国新世纪优秀人才支持计划(NCET-05-0774);重庆市自然科学基金(杰出青年科学基金)(CSTC;2008BA5040)

摘　　要：目的比较几种人偏肺病毒(hMPV)的检测方法,为临床选择适当的检测方法提供依据。方法不同稀释度的hMPV分别感染Vero-E6细胞,出现细胞病变(CPE)后,分别用RT-PCR、Real-time PCR和IFA进行检测,比较3种方法的灵敏度;10倍系列稀释滴度为105.2TCID50/ml的hMPV,分别用RT-PCR及Real-time PCR进行检测,定量比较两种方法的灵敏度;分别用Real-time PCR和IFA检测523份急性呼吸道感染患儿的鼻咽分泌物。结果IFA、RT-PCR和Real-time PCR分别可检出10-4、10-5、10-7稀释度孔中的病毒;RT-PCR和Real-time PCR检测的灵敏度分别为1.0×103和1TCID50/ml;在523份鼻咽分泌物标本中,IFA检测阳性率为9.18%,Real-time PCR检测阳性率为31.55%。结论Real-time PCR检测鼻咽分泌物标本中hMPV为儿科临床早期诊断的最佳方法;IFA可作为基层医院及hMPV相关严重呼吸道感染的检测方法。Objective To compare various methods for determination of human metapneumovirus（hMPV）and provide a basis for selection of an appropriate method used in clinic.Methods Vero-E6 cells were infected with hMPV at various dilutions and,after appearance of CPE,determined by RT-PCR,Real-time PCR and IFA respectively to compare the sensitivities of the three methods. The hMPV at a titer of 105.2 TCID50/ml was diluted 10-fold serially and determined by RT-PCR and Real-time PCR respectively to compare the sensitivities of the two methods quantitatively.The nasopharynx secretes of 523 children with acute respiratory infection were determined by Real-time PCR and IFA respectively.Results The hMPV at dilutions of 10-4,10-5 and 10-7 were detected by IFA,RT-PCR and Real-time PCR respectively.The sensitivities of RT-PCR and Real-time PCR were 1×103 and 1 TCID50/ml respectively.The positive rates of hMPV in 523 nasopharynx secrete specimens determined by IFA and Real-time PCR were 9.18% and 31.55%respectively.Conclusion Real-time PCR is the first choice for early clinical diagnosis of hMPV infection in nasopharynx secretes of children.IFA may be used for diagnosis of severe hMPV-associated respiratory infection and for the diagnosis in grassroot hospital.

关 键 词：人偏肺病毒 反转录聚合酶链反应 实时荧光定量聚合酶链反应 间接免疫荧光试验 

分 类 号：R373.9[医药卫生—病原生物学] R392.33[医药卫生—基础医学]