Abstract

The outbreak of the emerging SARS-CoV-2 virus has highlighted the challenges of detecting viral infections, especially in resource-limited settings. The SARS-CoV-2 virus transmission chain is interrupted when screening and diagnosis can be performed on a large scale by identifying asymptomatic or moderately symptomatic patients. Diagnosis of COVID-19 with reverse transcription polymerase chain reaction (RT-PCR) has been limited due to inadequate access to complex, expensive equipment and reagents, which has impeded efforts to reduce the spread of virus transmission. Recently, the development of several diagnostic platforms based on the CRISPR-Cas system has reduced the dependence on RT-PCR. The first CRISPR-based diagnostic test for SARS-CoV-2 was recently approved by the U.S. Food and Drug Administration. The biosensing systems have several important features that make them suitable for point-of-care tests, including the speed of design and synthesis of each platform in less than a few days, an assay time of 1–2 h, and the cost of materials and reagents less than one dollar per test. The HUDSON-SHERLOCK and STOPCovid biosensing systems, as field-deployable and rapid diagnostic tests, can detect low-copy viruses in body fluids without nucleic acid extraction and with minimal equipment. In addition, Cas13-based treatment strategies could potentially be an effective antiviral strategy for the prevention and treatment of emerging pandemic viruses such as SARS-CoV-2. In this review, we describe recent advances in CRISPR-based diagnostic platforms with an emphasis on their use in the rapid diagnosis and potential treatment of COVID-19.

摘要

新出现的 SARS-CoV-2 病毒的爆发凸显了检测病毒感染的挑战，尤其是在资源有限的环境中。当可以通过识别无症状或中度症状的患者进行大规模筛查和诊断时，SARS-CoV-2 病毒传播链被中断。由于无法获得复杂、昂贵的设备和试剂，因此通过逆转录聚合酶链反应 (RT-PCR) 诊断 COVID-19 受到限制，这阻碍了减少病毒传播的努力。最近，基于 CRISPR-Cas 系统的几种诊断平台的开发减少了对 RT-PCR 的依赖。美国食品和药物管理局最近批准了第一个基于 CRISPR 的 SARS-CoV-2 诊断测试。生物传感系统有几个重要的特点，使其适用于即时检验，包括在不到几天的时间内设计和合成每个平台的速度、1-2 小时的分析时间以及材料成本和试剂每次测试不到一美元。HUDSON-SHERLOCK 和 STOPCovid 生物传感系统作为可现场部署的快速诊断测试，无需核酸提取和最少的设备即可检测体液中的低拷贝病毒。此外，基于 Cas13 的治疗策略可能是预防和治疗 SARS-CoV-2 等新兴大流行病毒的有效抗病毒策略。在这篇综述中，我们描述了基于 CRISPR 的诊断平台的最新进展，重点是它们在 COVID-19 的快速诊断和潜在治疗中的应用。