Deployable CRISPR-Cas13a diagnostic tools to detect and report Ebola and Lassa virus cases in real-time
Deployable CRISPR-Cas13a diagnostic tools to detect and report Ebola and Lassa virus cases in real-time
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DOI：
10.1038/s41467-020-17994-9
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发表时间：
2020-08-17
影响因子：
16.6
通讯作者：
Sabeti, Pardis C.
中科院分区：
文献类型：
--
作者：
Barnes, Kayla G.;Lachenauer, Anna E.;Sabeti, Pardis C.
最近爆发的病毒性出血热（VHF），包括埃博拉病毒病（EVD）和拉沙热（LF），突出了对敏感的，可部署的测试来诊断这些毁灭性的人类疾病的迫切需要。在这里，我们开发了基于CRISPR-Cas 13 a（SHERPRTM）的诊断方法，针对埃博拉病毒（EBOV）和拉沙病毒（LASV），具有荧光和侧向流读数。我们在实验室和临床样本上证明了这些检测试剂的灵敏度以及SHERRENT平台应对病毒特异性诊断挑战的能力。我们进行安全性测试，以证明我们的哈德逊协议在SHERMOT测试之前热灭活VHF病毒的有效性，从而无需提取。我们开发了一个用户友好的协议和移动的应用程序（HandLens）来报告结果，促进SHERRisk在流行地区的使用。最后，我们成功地在塞拉利昂和尼日利亚部署了我们的测试，以应对最近的疫情。病毒性出血热的爆发突出了对灵敏的、可现场部署的诊断方法的需要。在本文中，作者介绍了一个基于CRISPR的SHERcycle平台，该平台具有用于埃博拉和拉沙热爆发的现场协议和移动的应用程序。
Recent outbreaks of viral hemorrhagic fevers (VHFs), including Ebola virus disease (EVD) and Lassa fever (LF), highlight the urgent need for sensitive, deployable tests to diagnose these devastating human diseases. Here we develop CRISPR-Cas13a-based (SHERLOCK) diagnostics targeting Ebola virus (EBOV) and Lassa virus (LASV), with both fluorescent and lateral flow readouts. We demonstrate on laboratory and clinical samples the sensitivity of these assays and the capacity of the SHERLOCK platform to handle virus-specific diagnostic challenges. We perform safety testing to demonstrate the efficacy of our HUDSON protocol in heat-inactivating VHF viruses before SHERLOCK testing, eliminating the need for an extraction. We develop a user-friendly protocol and mobile application (HandLens) to report results, facilitating SHERLOCK's use in endemic regions. Finally, we successfully deploy our tests in Sierra Leone and Nigeria in response to recent outbreaks. Outbreaks of viral hemorrhagic fevers highlight the need for sensitive, field-deployable diagnostics. Here the authors present a CRISPR-based SHERLOCK platform with field protocol and mobile app for Ebola and Lassa fever outbreaks.