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用逆转录PCR-核酸探针杂交法对柯萨奇B组病毒分型诊断
引用本文:李振勇,李智涛,冯艳铭,邵大晓,赵大鹏,崔天星,杨国翠,屈凌波,赵玉芬.用逆转录PCR-核酸探针杂交法对柯萨奇B组病毒分型诊断[J].中华实验和临床病毒学杂志,2004,18(3):291-293.
作者姓名:李振勇  李智涛  冯艳铭  邵大晓  赵大鹏  崔天星  杨国翠  屈凌波  赵玉芬
作者单位:1. 450052,郑州大学化学系化学生物实验室
2. 华美生物工程公司
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152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。

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
修稿时间:2004年1月2日

Coxsackie virus B types were discriminated by RT-PCR
LI Zhen-yong ,LI Zhi-tao,FENG Yan-ming,SHAO Da-xiao,ZHAO Da-peng,CUI Tian-xing,YANG Guo-cui,QU Ling-bo,ZHAO Yu-fen Laboratory of Chemobiology.Coxsackie virus B types were discriminated by RT-PCR[J].Chinese Journal of Experimental and Clinical Virology,2004,18(3):291-293.
Authors:LI Zhen-yong  LI Zhi-tao  FENG Yan-ming  SHAO Da-xiao  ZHAO Da-peng  CUI Tian-xing  YANG Guo-cui  QU Ling-bo  ZHAO Yu-fen Laboratory of Chemobiology
Institution:Laboratory of Chemobiology, Department of Chemistry, Zhengzhou University, Zhengzhou 450052, China. zhenyongl@sabc.com.cn
Abstract: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
Keywords:Polymerase chain reaction  Enterovirus B  human  Diagnosis  differential
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