乙型肝炎病毒（HBV）和丙型肝炎病毒（HDV）的共同感染导致了丙型肝炎，这是最严重的慢性病毒性肝炎形式，经常导致肝功能衰竭和肝细胞癌。长效干扰素α是多年来慢性丙型肝炎唯一的治疗选择，但疗效有限。新的治疗方法正在进行进一步的临床开发中，最近已经获得一项批准。丙型肝炎的诊断和抗病毒治疗效果监测基于HDV RNA的检测和定量。然而，可靠的HDV RNA检测方法的开发受到病毒的异质性（至少8种不同基因型和多个亚基因型）、宿主体内病毒的多样性、病毒快速进化以及HDV RNA独特的二级结构特征的挑战。不同的RNA提取方法、核酸检测试剂盒的引物/探针设计、自动化的缺乏以及检测实验室之间标准化的普遍缺乏都导致了研究和商业HDV RNA检测方法性能特征的显著差异。世界卫生组织（WHO）已经提供了大约10年的HDV RNA标准，许多实验室使用它来确定自己实验的检测限度，并促进研究中心之间RNA水平的比较。本文回顾了可靠的泛基因型HDV RNA定量的挑战，讨论了特定的临床需求和可靠的HDV RNA定量在药物开发和患者监测中的重要性。我们总结了可用的HDV RNA检测方法的不同技术特征和性能特征。最后，我们提供了在药物开发和患者监测中使用HDV RNA检测方法的考虑事项。版权所有© 2023美国肝病研究协会。

Co-infection with hepatitis B virus (HBV) and hepatitis D virus (HDV) results in hepatitis D, the most severe form of chronic viral hepatitis, frequently leading to liver decompensation and hepatocellular carcinoma. Pegylated interferon alpha, the only treatment option for chronic hepatitis D for many years, has limited efficacy. New treatments are in advanced clinical development, with one recent approval. Diagnosis and antiviral treatment response monitoring are based on detection and quantification of HDV RNA. However, the development of reliable HDV RNA assays is challenged by viral heterogeneity (at least 8 different genotypes and several subgenotypes), intra-host viral diversity, rapid viral evolution, and distinct secondary structure features of HDV RNA. Different RNA extraction methodologies, primer/probe design for Nucleic Acid Tests, lack of automation, and overall dearth of standardization across testing laboratories contribute to substantial variability in performance characteristics of research-based and commercial HDV RNA assays. A WHO standard for HDV RNA, available for about 10 years, has been used by many laboratories to determine the limit of detection of their assays and facilitates comparisons of RNA levels across study centers. Here we review challenges for robust pan genotype HDV RNA quantification, discuss particular clinical needs and the importance of reliable HDV RNA quantification in the context of drug development and patient monitoring. We summarize distinct technical features and performance characteristics of available HDV RNA assays. Finally, we provide considerations for the use of HDV RNA assays in the context of drug development and patient monitoring.Copyright © 2023 American Association for the Study of Liver Diseases.