半巢式RT-PCR法检测呼吸道合胞病毒

Detection of respiratory syncytial virus by semi-nested RT-PCR

目的 建立快速、敏感、特异的人呼吸道合胞病毒 (Respiratorysyncytialvirus ,RSV)检测方法。 方法 根据RSVF基因的保守序列设计 3条引物 ,建立半巢式RT -PCR(semi-nestedRT -PCR)检测方法。用该方法检测12 5例患急性下呼吸道感染的婴幼儿的鼻咽吸引物 (nasopharyngealaspirates,NPA)标本 ,同时对标本进行病毒分离和直接免疫荧光法检测。结果 该半巢式RT -PCR灵敏度为 0 .1TCID50 ,用该方法在 12 5例标本中共检出RSV阳性标本 6 3例 ,阳性率 5 0 .4 %。结论 建立快速、敏感、特异的RSV半巢式RT -PCR检测方法 。

Objective To develop a rapid,sensitive,speci fic method to detect human respiratory syncytial virus(RSV).Methods Three pieces of primer were desig ned from the conservative F gene of RSV and a semi-nested RT-PCR method was achi eved. This method was used to diagnose 125 nasopharyngeal aspirates obtained fro m children with lower respiratory tract infection,at the same time the specimen s were also analyzed by viral isolation and direct fluorescent-antibody assay (D FA).Results The sensitivity of the semi-nested RT-PCR assay was determined to be 0.1TCID 50; 63 cases (50.4%)were identified RSV-positi ve by the semi-nested RT-PCR. Conclusion The semi-nested RT-PCR was a rapid,sensitive ,specific method for diagnosis of RSV infection,and it could be used for early c linical diagnosis as well as epidemiological surveillance.