Public Health England, National Infection Service, Porton Down, Salisbury, Wiltshire, United Kingdom.
PLoS Negl Trop Dis. 2020 Jul 31;14(7):e0008496. doi: 10.1371/journal.pntd.0008496. eCollection 2020 Jul.
The unprecedented 2013/16 outbreak of Zaire ebolavirus (Ebola virus) in West Africa has highighted the need for rapid, high-throughput and POC diagnostic assays to enable timely detection and appropriate triaging of Ebola Virus Disease (EVD) patients. Ebola virus is highly infectious and prompt diagnosis and triage is crucial in preventing further spread within community and healthcare settings. Moreover, due to the ecology of Ebola virus it is important that newly developed diagnostic assays are suitable for use in both the healthcare environment and low resource rural locations.
METHODOLOGY/PRINCIPLE FINDINGS: A LAMP assay was successfully developed with three detection formats; a real-time intercalating dye-based assay, a real-time probe-based assay to enable multiplexing and an end-point colourimetric assay to simplify interpretation for the field. All assay formats were sensitive and specific, detecting a range of Ebola virus strains isolated in 1976-2014; with Probit analysis predicting limits of detection of 243, 290 and 75 copies/reaction respectively and no cross-detection of related strains or other viral haemorrhagic fevers (VHF's). The assays are rapid, (as fast as 5-7.25 mins for real-time formats) and robust, detecting Ebola virus RNA in presence of minimally diluted bodily fluids. Moreover, when tested on patient samples from the 2013/16 outbreak, there were no false positives and 93-96% of all new case positives were detected, with only a failure to detect very low copy number samples.
CONCLUSION/SIGNIFICANCE: These are a set of robust and adaptable diagnostic solutions, which are fast, easy-to-perform-and-interpret and are suitable for use on a range of platforms including portable low-power devices. They can be readily transferred to field-laboratory settings, with no specific equipment needs and are therefore ideally placed for use in locations with limited resources.
2013-2016 年扎伊尔埃博拉病毒（埃博拉病毒）在西非的空前爆发凸显了对快速、高通量和 POCT 诊断检测的需求，以实现对埃博拉病毒病（EVD）患者的及时检测和适当分诊。埃博拉病毒具有高度传染性，及时诊断和分诊对于防止在社区和医疗环境中进一步传播至关重要。此外，由于埃博拉病毒的生态，新开发的诊断检测方法必须适合在医疗环境和资源匮乏的农村地区使用。
方法/原理发现：成功开发了一种 LAMP 检测法，具有三种检测形式：实时嵌入染料检测法、实时探针检测法以实现多重检测和终点比色检测法以简化现场解释。所有检测方法都具有敏感性和特异性，可检测 1976-2014 年分离的多种埃博拉病毒株；概率分析预测检测限分别为 243、290 和 75 拷贝/反应，并且没有交叉检测到相关株或其他病毒性出血热（VHF）。这些检测方法快速（实时格式最快为 5-7.25 分钟）且稳健，可在最小稀释体液存在下检测到埃博拉病毒 RNA。此外，在对 2013-2016 年爆发期间的患者样本进行测试时，没有假阳性，93-96%的新病例阳性都被检测到，只有在检测到非常低拷贝数样本时才会失败。
结论/意义：这些是一组稳健且适应性强的诊断解决方案，快速、易于操作和解释，适用于多种平台，包括便携式低功率设备。它们可以很容易地转移到现场实验室环境中，无需特定设备，因此非常适合在资源有限的地方使用。
