Department of Clinical and Toxicological Analyses, Faculty of Pharmacy, Federal University of Minas Gerais, Belo Horizonte, Minas Gerais, Brazil.
Department of Genetics and Morphology, Institute of Biological Sciences, University of Brasilia, Brasilia 70910-900, Brazil.
Biomed Pharmacother. 2022 Sep;153:113538. doi: 10.1016/j.biopha.2022.113538. Epub 2022 Aug 12.
The World Health Organizations declaration of the COVID-19 pandemic was a milestone for the scientific community. The high transmission rate and the huge number of deaths, along with the lack of knowledge about the virus and the evolution of the disease, stimulated a relentless search for diagnostic tests, treatments, and vaccines. The main challenges were the differential diagnosis of COVID-19 and the development of specific, rapid, and sensitive tests that could reach all people. RT-PCR remains the gold standard for diagnosing COVID-19. However, new methods, such as other molecular techniques and immunoassays emerged. Also, the need for accessible tests with quick results boosted the development of point of care tests (POCT) that are fast, and automated, with high precision and accuracy. This assay reduces the dependence on laboratory conditions and mass testing of the population, dispersing the pressure regarding screening and detection. This review summarizes the advances in the diagnostic field since the pandemic started, emphasizing various laboratory techniques for detecting COVID-19. We reviewed the main existing diagnostic methods, as well as POCT under development, starting with RT-PCR detection, but also exploring other nucleic acid techniques, such as digital PCR, loop-mediated isothermal amplification-based assay (RT-LAMP), clustered regularly interspaced short palindromic repeats (CRISPR), and next-generation sequencing (NGS), and immunoassay tests, and nanoparticle-based biosensors, developed as portable instruments for the rapid standard diagnosis of COVID-19.
世界卫生组织宣布 COVID-19 大流行是科学界的一个里程碑。高传播率和大量死亡，以及对病毒和疾病演变的知识缺乏，刺激了对诊断测试、治疗方法和疫苗的不懈探索。主要挑战是 COVID-19 的鉴别诊断和开发能够覆盖所有人的特定、快速和敏感的测试。实时聚合酶链反应（RT-PCR）仍然是诊断 COVID-19 的金标准。然而，新的方法，如其他分子技术和免疫测定法已经出现。此外，需要具有快速结果的易于使用的测试，推动了即时检测（POCT）的发展，这些测试快速、自动化，具有高精度和准确性。该检测方法减少了对实验室条件的依赖和对人群的大规模检测，缓解了筛查和检测方面的压力。这篇综述总结了大流行开始以来诊断领域的进展，强调了用于检测 COVID-19 的各种实验室技术。我们回顾了主要的现有诊断方法，以及正在开发的 POCT，从 RT-PCR 检测开始，但也探索了其他核酸技术，如数字 PCR、环介导等温扩增检测（RT-LAMP）、簇状规律间隔短回文重复序列（CRISPR）和下一代测序（NGS），以及免疫测定测试和基于纳米颗粒的生物传感器，作为 COVID-19 快速标准诊断的便携式仪器进行开发。
