Raw data for Tables S2-S4.

Isothermal nucleic acid amplification methods, such as recombinase polymerase amplification (RPA), are becoming increasingly vital as diagnostic platforms for neglected tropical diseases by enabling rapid and accurate detection in under-resourced regions. We developed a real-time RT-RPA assay for Machupo virus (MACV), the causative agent of Bolivian hemorrhagic fever, and directly compared it to a real-time RT-PCR assay targeting the same viral sequence fragment. The methods were evaluated across critical parameters: limit of detection (LOD), tolerance to single-nucleotide substitutions, multiplexing capability, and adaptability to multiple MACV genetic variants. The LOD was identical for both assays: 5 × 103 copies/ml of armored RNA particles. They differed in terms of input RNA (copies/reaction): 100 (PCR) versus 20 (RPA). The real-time RT-RPA assay was further validated on a portable device, demonstrating its potential for field-deployability for point-of-care applications.恒温核酸扩增技术（isothermal nucleic acid amplification methods），如重组酶聚合酶扩增（recombinase polymerase amplification, RPA），正日益成为被忽视热带病的重要诊断平台，可在资源匮乏地区实现快速精准的病毒检测。本研究针对玻利维亚出血热的病原体马秋波病毒（Machupo virus, MACV）开发了实时逆转录RPA检测方法，并与靶向同一病毒序列片段的实时逆转录PCR检测方法进行了直接比对。本研究从以下关键参数对两种方法开展评估：检测限（limit of detection, LOD）、单核苷酸替换耐受性、多重检测能力以及对多种MACV遗传变异株的适配性。两种检测方法的LOD完全一致：均为5×10³拷贝/毫升的装甲RNA颗粒（armored RNA particles）。二者在输入RNA量（拷贝数/反应体系）上存在差异：实时逆转录PCR为100拷贝/反应，实时逆转录RPA为20拷贝/反应。本研究进一步在便携式设备上对该实时逆转录RPA检测方法进行了验证，证实其具备现场即时检测的应用潜力。