实时荧光PCR检测丙型肝炎病毒核酸及其临床意义

Dectection of the Hepatitis C Virus Nucleoside with Real - time Fluorescence PCR and it's Clinical Significance

目的:探讨实时荧光PCR检测技术在丙型肝炎病原诊断及在监测干扰素-α治疗慢性丙型肝炎早期病毒学应答反应(EVR)中的临床意义。方法:采用一种新的丙型肝炎病毒核酸(HCV-RNA)扩增(PCR)定量检测方法——实时荧光PCR检测HCV-RNA,对96％例慢性丙肝(其中20例随机接受Pegasys或Referon-α治疗,每例患者分别采集治疗前后系列血清7份)、30例其它肝病、15例非肝病和86例健康献血员的血清标本进行HCV-RNA检测,部分血清标本与Am-plicor(Roche)进行核对。结果:96例慢性丙肝患者血清HCV-RNA检出率为85.4％(82/96)、其它肝病、非肝病和健康献血员中均未检出HCV-RNA;部分标本经与Amplicor核对呈较好相关性,r(log)=0.91。随机接受Pegasys或Referon-α治疗的20例丙肝患者治疗前后EVR显示,前者的EVR似较后者明显,但因病例数太少,无明显统计学差异(G=4.467,P>0.05)。结论:实时荧光PCR检出的HCV-RNA载量可较客观地反映丙肝患者体内病毒复制水平,支持干扰素治疗慢性丙型肝炎EVR可用于预测长期疗效。

Objective: To explore the clinical significance of real - time fluorescence RT- PCR quantification of HCV - RNA in the clinical diagnosis of chronic hepatitis C and in evaluating the effects of antiviral treatment. Metbods: Real - time fluorescence RT - PCR was used to detect 96 patients vrith chronic hepatitis C (randomly assigned 20 of them to treated with Pegasys or Referon -a), 30 patients suffering from other liver disease, 15 patients suffering from other disease, and 86 healthy donors, Amplicor HCV Moni-torTM Test, as control, was used to detect some of the serum specimens. Results: The positive detection rate of HCV - RNA in 96 patients with hepatitis C was 85.4% ( 82/96), HCV - RNA were not detected in samples from 30 patients suffering from other hepatic diseases, 15 patients suffering from other disease and 86 donors. Significant correlation was found between the results of real - time quan-titative PCR and Amplicor in Some of the samples (r = 0.91). In 20 patients with chronic hepatitis C who received Pegasys or Referon - atherapy, ealer virologic response (EVR)was observed in the Pegasys groups more than in Referon - agroups, but with no statis-tics difference(G = 4.467, P > 0.05) . Conclusion: The HCV - RNA detected with quantification real - time fluorescence PCR can show the HCV replication Ievel in patients. Our results also suggested that the EVR may be used to predict the IFN therapy sustained response in chronic hepatitis C.