呼吸道合胞病毒检测方法比较

Comparison of four methods used in detection of the respiratory syncytial virus In nasopharyngeal aspirates from children with the acute lower respiratory tRact infection

为使临床治疗更及时有效 ,以通过不同方法的比较得到更快速、敏感、特异的早期从临床标本中检测呼吸道合胞病毒的方法。对收集的80份疑为病毒性急性下呼吸道感染患儿鼻咽分泌物标本应用不同方法进行了RSV检测 ,包括病毒分离、间接免疫荧光、巢式PCR及核酸杂交。80份标本都进行了病毒分离及巢式PCR ;80份标本沉淀分为两组 ,第1组32份 (32/80份 )进行间接免疫荧光 ;第2组48份 (48/80份 )进行杂交。结果显示 ,80份标本中除外9份在病毒分离过程中出现污染 ,RSV分离组阳性标本22份 (22/71份 ,31.0% ) ;间接免疫荧光组阳性标本14份 (14/32份 ,43.8 % ) ;核酸杂交组阳性标本22份 (22/48 ,45.8 % ) ,其中A亚型13份 ,B亚型9份 ;巢式PCR检测方法中共有66份 (66/80份 ,82.5 % )有目的基因片段扩增 ,A亚型45份 ,B亚型21份。结果表明 ,从临床标本中检测RSV以巢式PCR灵敏度最高 ;杂交法与免疫荧光法阳性率大致相同 ;病毒分离特异性好 ,但由于其他原因导致了分离率低。

in order to select more rapid,sensitive and specific method in dEtEction of rsv directly from the clinical specimens,rsv were detected with nesteD_pcr,dot blot hybridization and virus isolation in tissue culture as well as iNdirect immunofluorescence assay(ifa)from nasopharyngeal aspirates collected fRom infants and young children with acute lower respiratory tract infections who were admitted into the wards.all of80specimens were tested for rsv with tissue CUlture and nested_pcr.in addition,32of the80specimens were tested with ifa and THe remaining48specimens with dot blot hybridization.the results showed rsv were ISolated from22(22/71,31.0%)specimcns out of80,with9specimens disˉcarded becAusc of contamination.fourteen(14/32,43.8%)were considered as rsv positive wiTh ifa and22(22/48,45.8%)were diagnosed as rsv positive with dot blot hybridiZation.the targeting gene fragment amplifications could be found in66specimens(66/80,82.5%)with nested_pcr assay(45of them belonged to a subset,21belonged TO b subset)indicating the nested_pcr is the most sensitive method in detection OF rsv from clinical specimens.ifa and dot blot hybridization methods have similaR sensitivity by comparing the positive rates from different methods.the tissue CUlture in detecting rsv is specific,but with lower sensitivity because of varioUs reasons.while the nested_pcr and dot blot hybridization have the advantage of direct determination of the rsv subset from the clinical samples.