实时逆转录PCR快速鉴别呼吸道合胞病毒亚型  

Rapid identification of subtypes of respiratory syncytial virus by real-time PCR

作　　者：林虹[1] 郑晓群[2] 

机构地区：[1]温州医学院检验医学院2000年级,浙江温州325035 [2]温州医学院附属第二医院检验科,浙江温州325027

出　　处：《检验医学教育》2005年第4期34-36,共3页

摘　　要：目的建立SYBR Green实时RT-PCR快速检测呼吸道合胞病毒(respiratory syncytial virus,RSV)A、B亚型的方法。方法根据RSV G蛋白编码基因的核苷酸序列设计三条引物,其中P1为RSV通用引物,P2、P3分别为A、B亚型特异性引物。使用SYBR Green荧光染料结合熔解曲线分析进行检测,根据产物特征性熔解温度(melting temperatures Tm)鉴别RSV A、B亚型。结果本法对RSV Long株(A亚型)和CH18375株(B亚型)进行检测,其Tm值分别为84.06和89.06℃;与常见呼吸道病毒间无交叉反应;48例临床标本中检出RSV阳性29例,其中A亚型占72.4%(21/29),B亚型占27.6%(8/29)。结论SYBR Green实时RT-PCR结合熔解曲线分析检测RSV亚型具有快速、简便、特异等特点,可对临床标本直接分型。Objective：To establish a rapid assay for detection of respiratory syncytial virus （RSV） A and B subtype. Methods： According to RSV G protein sequence, three primers which contained one primer for both subtypes and two for either subtype A or B were designed. Detection of RSV A and B were based on determination of specific melting temperatures by real-time PCR and melting curve analysis using SYBR Green Ⅰ fluorescent dye. Results：Tm value of RSV Long strains （subtype A） and CH18375 strains（subtype B） by using this method was 84. 06 the other respiratoty viruses was observed. Of the 48 samples, and 89.06℃ ,respectively ;there was no cross reactivity with 29 were positive for RSV,and 21 （72. 4%） of them were subtype A, 16 （27.6%） of them were subtype B. Conclusions：Detection RSV subtype by real-time RT-PCR and melting curve analysis using SYBR Green Ⅰ is more rapid, simple and specificity.

关 键 词：呼吸道合胞病毒 实时PCR 熔解曲线分析 

分 类 号：R735.2[医药卫生—肿瘤] R373.1[医药卫生—临床医学]