荧光定量两探针法检测HCV RNA的研究

Study on fluorescence quantitative RT-PCR with two-probes for detection of HCV RNA

目的建立荧光定量两探针检测HCVRNA方法。方法对105例抗HCV抗体阳性和220例阴性样本,用荧光定量两探针法和2种商品荧光定量试剂进行检测,并对检测结果进行比较。结果荧光定量两探针法阳性率分别为89.5((94/105)和2.3((5/220)。两种商品荧光定量试剂阳性率分别为71.4((75/105)和0.45((1/220);69.5((73/105)和1.8((4/220)。结论荧光定量两探针法检测HCVRNA有较高的敏感性和特异性。

Objective To establish a fluorescence quantitative-reverse transcription-polymerase chain reaction （FQ-RT-PCR） with twoprobe for quantification of HCV RNA loads, and to optimize the experimental conditions to increase sensitivity and specificity. Methods HCV RNA loads in 89 cases with positive anti-HCV and 220 cases with negative anti-HCV were quantified by FQ-RT-PCR with double probe,and the results were compared with another two commercial HCV RNA quantificationkits simultaneously. Results For the 2 groups （89 cases with positive anti-HCV and 220 cases with negative anti-HCV）, the positive rate of HCV RNA was 91.0 % （81 cases） and was 2.27% （5 cases） respectively by FQ-RT-PCR with two-probe, while it was 82.0 % （73 cases）and 0.45 % （1 cases） ）, 79.7 % （71 cases） ） and 1.81% （4 cases） respectively by using the two commercial kits. Conclusion FQ-RT-PCR assay with two-probe may increases the sensitivity and specificity for the detection of HCV RNA loads comparing with commercial kits.