Department of Biochemistry, University of Washington, Seattle, Washington, United States of America.
Department of Mechanical Engineering, University of Washington, Seattle, Washington, United States of America.
PLoS One. 2022 Oct 25;17(10):e0276582. doi: 10.1371/journal.pone.0276582. eCollection 2022.
Over 71 million people are infected with hepatitis C virus (HCV) worldwide, and approximately 400,000 global deaths result from complications of untreated chronic HCV. Pan-genomic direct-acting antivirals (DAAs) have recently become widely available and feature high cure rates in less than 12 weeks of treatment. The rollout of DAAs is reliant on diagnostic tests for HCV RNA to identify eligible patients with viremic HCV infections. Current PCR-based HCV RNA assays are restricted to well-resourced central laboratories, and there remains a prevailing clinical need for expanded access to decentralized HCV RNA testing to provide rapid chronic HCV diagnosis and linkage to DAAs in outpatient clinics. This paper reports a rapid, highly accurate, and minimally instrumented assay for HCV RNA detection using reverse transcription recombinase polymerase amplification (RT-RPA). The assay detects all HCV genotypes with a limit of detection of 25 copies per reaction for genotype 1, the most prevalent in the United States and worldwide. The clinical sensitivity and specificity of the RT-RPA assay were both 100% when evaluated using 78 diverse clinical serum specimens. The accuracy, short runtime, and low heating demands of RT-RPA may enable implementation in a point-of-care HCV test to expand global access to effective treatment via rapid chronic HCV diagnosis.
全球有超过 7100 万人感染丙型肝炎病毒（HCV），未经治疗的慢性 HCV 感染并发症导致约 40 万人死亡。全基因组直接作用抗病毒药物（DAAs）最近已广泛应用，在不到 12 周的治疗时间内，治愈率很高。DAA 的推出依赖于 HCV RNA 的诊断检测，以确定有资格接受病毒血症 HCV 感染治疗的患者。目前基于 PCR 的 HCV RNA 检测方法仅限于资源充足的中心实验室，临床仍需要扩大去中心化的 HCV RNA 检测，以提供快速的慢性 HCV 诊断，并在门诊诊所中与 DAA 联系。本文报告了一种使用逆转录重组酶聚合酶扩增（RT-RPA）检测 HCV RNA 的快速、高度准确且仪器要求较低的方法。该检测方法可检测所有 HCV 基因型，对美国和全球最常见的 HCV 基因型 1 的检测限为每个反应 25 个拷贝。使用 78 种不同的临床血清标本评估时，RT-RPA 检测的临床灵敏度和特异性均为 100%。RT-RPA 的准确性、短运行时间和低加热要求可能使其能够在即时护理 HCV 检测中实施，从而通过快速慢性 HCV 诊断扩大有效治疗的全球可及性。
