用逆转录PCR-核酸探针杂交法对柯萨奇B组病毒分型诊断  

Coxsackie virus B types were discriminated by RT-PCR

作　　者：李振勇[1] 李智涛 冯艳铭 邵大晓 赵大鹏 崔天星 杨国翠 屈凌波[1] 赵玉芬[1] 

机构地区：[1]郑州大学化学系化学生物实验室,450052 [2]华美生物工程公司

出　　处：《中华实验和临床病毒学杂志》2004年第3期291-293,共3页Chinese Journal of Experimental and Clinical Virology

摘　　要：目的　建立一种逆转录PCR 核酸探针杂交法对柯萨奇B组病毒 (CVB1～ 6 )进行分型诊断。方法　一对通用PCR引物 ,它能有效扩增所有CVB1～ 6型的特异性DNA片段 ;另选取 6条各型特异性寡核苷酸探针 ,将它们分别共价结合在不同的微孔板上。经过一次PCR扩增 ,扩增后的产物分别与包被有不同探针的微孔板进行杂交检测 ,从而有效鉴别CVB各型。结果　本法与ELISA法的分型比较显示它们具有很好的一致性 ,无错误分型。对 15 2例IgM抗体阳性标本的检测 ,该方法阳性率为 71 7%。结论　本法可准确对CVB进行分型 ,为CVB的临床诊断及流行病学调查提供了一种有效的方法。Objective To develop a method for detection of coxsackie B virus type 1-6 by RT-PCR Methods A pair of primers were designed to amplify all types of coxsackie B virus 1-6 efficiently The PCR product was hybridized in micro-wells in which 6 type specific oligonucleotide probes had been coated respectively, colorimetric detection was performed to discriminate the types of coxsackie B virus Results This method was shown to be concordant with the IgM ELISA, 717% of anti-coxsackie B positive cases could be detected by RT-PCRConclusion The RT-PCR method can type coxsackie B virus efficiently and provides a tool for clinical diagnosis and epidemiological investigation

关 键 词：CVB 柯萨奇B组病毒 逆转录PCR 分型诊断 微孔板 杂交法 扩增 核酸探针 包被 PCR引物 

分 类 号：R446.5[医药卫生—诊断学]