急性呼吸道感染患儿人冠状病毒NL63检测与分析  

Detection and analysis of HCoV-NL63 in the children with acute respiratory tract infections

作　　者：陈云欢[1] 伍严安[1] 

机构地区：[1]福建医科大学省立临床医学院福建省立医院检验科,福州350001

出　　处：《中国实用儿科杂志》2010年第12期916-919,共4页Chinese Journal of Practical Pediatrics

基　　金：福建省自然科学基金资助项目(项目编号:F0310042)

摘　　要：目的建立人冠状病毒NL63(human coronavirus NL63)的实时荧光定量PCR(real-time fluorescentquantitative PCR,FQ-PCR)方法,了解呼吸道疾病患儿感染人冠状病毒NL63(HCoV-NL63)的情况。方法收集2008年10月至2009年4月因急性呼吸道感染(ARTI)而就诊于福建医科大学省立临床医学院患儿的咽拭子、鼻咽抽吸物、痰标本共151份,设计多聚酶蛋白1a基因的引物和Taqman探针,扩增1a基因片段,并将其克隆到PMD18-T载体上,构建质粒标准品,建立FQ-PCR检测方法,进行敏感性、特异性试验。扩增核衣壳蛋白N基因,对其序列进行初步分析。结果所建立的FQ-PCR方法特异性好,线性范围为101～1010copies/μL,变异系数小于5%。151份临床标本中共检测到2份HCoV-NL63阳性,阳性率为1.3%(2/151)。结论采用FQ-PCR方法可以检测急性呼吸道疾病患儿感染HCoV-NL63的情况。Objective To establish a real-time fluorescent quantitative PCR assay to detect the human coronavirus NL63 from nasopharyngeal samples of children with acute respiratory tract infections in Fuzhou.Methods The specific primers and Tap-man probes were designed targeting the 1a gene.The aimed fragment of 1a gene was amplified with PCR and ligated into a PMD18-T Easy vector for standards.A total of 151 clinical specimens were subsequently tested after determination of the sensitivity and specificity of the established real-time PCR.Amplify and preliminarily analyse the N gene.Results The specificity of this assay was excellent.The linear amplification of the assay ranged from 101 copies/μL to 1010 copies/μL.Two of 151 clinical specimens（1.3%） were tested positive for HCoV-NL63.Conclusion The real-time fluorescent quantitative PCR assay is successfully established to detect HCoV- NL63.

关 键 词：人冠状病毒NL63(HCoV-NL63) 急性呼吸道感染(ARTI) 实时荧光定量PCR N基因 

分 类 号：R72[医药卫生—儿科]