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丙型肝炎病毒(HCV)荧光PCR(F-PCR)试剂盒的研制及与免疫学方法的比较 被引量:10

Development of hepatitis C virus(HCV) fluorescence PCR (F-PCR) diagnostic kit and its clinical trial comparing with immunological method
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摘要 目的:用荧光PCR(Fluorescence PCR,F-PCR)方法研制丙型肝炎病毒(Hepatitis C Virus,HCV)RNA检测试剂盒,通过临床验证,考核其性能,并与免疫学方法比较。方法:F-PCR是PCR和荧光探针杂交技术结合所产生的PCR方法。由于采用完全闭管荧光检测,避免了PCR后处理导致的假阳性污染;又采用了荧光检测技术,可以提高检测的灵敏度。结果:设计合成了HLV F-PCR诊断试剂盒。检测了 512份临床血清标本,以美国 Abbott公司的HCV ELISA诊断试剂盒和美国Biotronics公司的HCV荧光RT-PCR诊断试剂盒(B-PCR)为对照。阳性率30.5%,灵敏度97.3%,特异性98.1%。结论:F-PC的灵敏度显著优于ELISA,也高于B-PCR;三者的特异性无统计学差异。F-PCR试剂盒可以检测HCV的真实感染情况,对于临床诊断和疗效考察有一定的指导意义。 Objective:Use a new F-PCR method to develop a hepatitis C virus(HCV) diagnostic kit, test the kit through clinical trial and compare it with immunological method. Methods: Fluorescence PCR(F-PCR) is a method which combines PCR and fluorescence probe hybridization together to measure DNA/RNA. Because in-tube monitoring of fluorescence signal can be done to stand for the quantitity of PCR product. Electrophoresis and UV detection are eliminated, so after-PCR cross-contamination which causes false positive can be avoided. Results:A clinical diagnostic kit for HCV with this method is developed. 512 clinical serum samples were tested with this kit, using HCV FLISA kit from Abbott Co.and HCV Fluorescence RT-PCR kit from Biotronics Co. (B-PCR) as control. The results shows :positive rate is 30.5%,sensitivity 97.3 % and specificity 98.1% . Conclusion: F-PCR is obviously superior to ELBA, and higher than B-PCR in sensitivity. The specificity of those three kits have no statistic difference. F-PCR can be used to monitor RNA of HCV in serum, and could be useful for clinical diagnose and therapy effects monitoring.
出处 《中国免疫学杂志》 CAS CSCD 北大核心 2002年第7期464-468,共5页 Chinese Journal of Immunology
基金 本研究为国家"863"中试项目(国科生字[2000]142号文) 2001年国家高新技术示范工程项目(计高科[2001]1214号文)部分工作 该试剂盒现已被批准为国家二类新生物制品(国药证字(200)S-44) 生产批准文号-国药试字S20000007
关键词 荧光PCR 丙型肝炎病毒 诊断试剂盒 HCV 丙型肝炎 诊断 免疫学诊断 荧光聚合酶链反应 Fluorescence PCR(F-PCR) Hepatitis C virus(HCV) Diagnostic kit
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参考文献5

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