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

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,春性能，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HCV 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。

Development of hepatitis C virus(HCV) fluorescence PCR (F-PCR) diagnostic kit and its clinical trial comparing with immunological method

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.