陈鹏 孙玉良 邓星

[摘要] 目的 評估高灵敏度实时荧光定量聚合酶链反应（real-time PCR）在病毒性肝病病毒核酸定量检测中的临床应用价值，为该方法的普及应用提供依据。 方法 收集2016年1月～2018年12月于湖北省中医院（以下简称“我院”）就诊的病毒性肝病患者895例，同期选取我院100名健康体检者作为对照组。分别应用常规方法和高灵敏度real-time PCR方法检测血清病毒核酸载量，比较两种方法检测结果的差异。对其中低病毒载量患者的生物化学指标、免疫学指标、凝血国际标准化比值（INR）和肝纤维化FIB-4指数的变化趋势、特点及其与病毒核酸载量的相关性进行统计分析。 结果 受检人群中低病毒载量患者呈高比例分布。高灵敏度real-time PCR方法检测HBV DNA的阳性率远高于常规方法（P < 0.05），而对于HCV RNA，两种方法检测结果的一致性较好。低病毒载量患者的多项生物化学指标波动并不明显，且多项生物化学指标与病毒核酸载量无相关性（P > 0.05）。大部分低病毒载量乙肝患者表现为HBeAg阴性的低水平复制状态。免疫学诊断指标与病毒核酸载量间存在不同程度的不一致和弱关联度。低病毒载量患者的凝血INR和肝纤维化FIB-4指数均高于对照组，差异有统计学意义（P < 0.05），但其与病毒核酸载量无相关性（P > 0.05）。 结论 高灵敏度real-time PCR方法对病毒核酸载量的检测具有不可比拟的优势，结合其他的诊断指标，其可在低拷贝病毒性肝病的病程管理中发挥不可替代的重要作用。

[关键词] 高灵敏度;实时荧光定量聚合酶链反应;病毒性肝病;病毒核酸定量;低病毒载量

[中图分类号] R446          [文献标识码] A          [文章编号] 1673-7210（2020）07（a）-0151-05

[Abstract] Objective To evaluate the clinical application value of high-sensitivity real-time polymerase chain reaction （real-time PCR） in the quantitative detection of viral nucleic acid in viral hepatitis， and to provide the basis for the widespread application of this method. Methods A total of 895 patients with viral hepatitis were enrolled from January 2016 to December 2018 in Hubei Provincial Hospital of Traditional Chinese Medicine （hereinafter referred to as “our hospital”）， and meanwhile another 100 normal people in our hospital were recruited as control group. Serum viral loads were detected by routine and high sensitive real-time PCR， then the difference of results between the two methods was compared. The trends and characteristics of biochemical indices，immunological markers， international standardized ratio of coagulation （INR）， index of liver fibrosis FIB-4 and their correlation with viral load were analyzed statistically in viral hepatitis patients with low viral load. Results The proportion of patients with low viral load was high among the population surveyed. The positive rate of HBV DNA detected by real-time PCR with high sensitivity was much higher than that by routine method（P < 0.05），but for HCV RNA， the two methods showed good consistency. The fluctuation of several biochemical indices was not obvious in low viral load patients， and multiple biochemical indexes were not correlated with viral nucleic acid load （P > 0.05）. Most hepatitis B patients with low viral load showed a status of low level HBeAg-negative replication. There were different degrees of inconsistency and weak correlation between immunological diagnostic markers and viral load results. INR of coagulation and FIB-4 index of liver fibrosis in patients with low viral load were higher than those in the control group， and the difference was statistically significant （P < 0.05）， but there was no correlation between them and viral nucleic acid load （P > 0.05）. Conclusion High sensitivity real-time PCR has incomparable advantages in the detection of viral nucleic acid，combined with other diagnostic markers，it can play an irreplaceable role in the course management of low viral load hepatitis.

[Key words] High sensitivity; Real-time polymerase chain reaction; Viral hepatitis; Viral nucleic acid quantification; Low viral load

由乙型肝炎病毒（HBV）和/或丙型肝炎病毒（HCV）引起的病毒性肝炎及其并發症是世界性的公共卫生问题，根据精准化的检测结果进行规范化的抗病毒治疗可有效地改善患者的生活质量[1-2]。……