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基于核酸扩增的 SARS-CoV-2 诊断:从基本概念到应用及其他。

SARS-CoV-2 Diagnostics Based on Nucleic Acids Amplification: From Fundamental Concepts to Applications and Beyond.

机构信息

National Institute for Agrarian and Veterinarian Research (INIAV, I.P), Vairão, Portugal.

Laboratory for Process Engineering, Environment, Biotechnology and Energy (LEPABE), Faculty of Engineering, University of Porto, Porto, Portugal.

出版信息

Front Cell Infect Microbiol. 2022 Mar 23;12:799678. doi: 10.3389/fcimb.2022.799678. eCollection 2022.

Abstract

COVID-19 pandemic ignited the development of countless molecular methods for the diagnosis of SARS-CoV-2 based either on nucleic acid, or protein analysis, with the first establishing as the most used for routine diagnosis. The methods trusted for day to day analysis of nucleic acids rely on amplification, in order to enable specific SARS-CoV-2 RNA detection. This review aims to compile the state-of-the-art in the field of nucleic acid amplification tests (NAATs) used for SARS-CoV-2 detection, either at the clinic level, or at the Point-Of-Care (POC), thus focusing on isothermal and non-isothermal amplification-based diagnostics, while looking carefully at the concerning virology aspects, steps and instruments a test can involve. Following a theme contextualization in introduction, topics about fundamental knowledge on underlying virology aspects, collection and processing of clinical samples pave the way for a detailed assessment of the amplification and detection technologies. In order to address such themes, nucleic acid amplification methods, the different types of molecular reactions used for DNA detection, as well as the instruments requested for executing such routes of analysis are discussed in the subsequent sections. The benchmark of paradigmatic commercial tests further contributes toward discussion, building on technical aspects addressed in the previous sections and other additional information supplied in that part. The last lines are reserved for looking ahead to the future of NAATs and its importance in tackling this pandemic and other identical upcoming challenges.

摘要

COVID-19 大流行引发了无数基于核酸或蛋白质分析的 SARS-CoV-2 诊断分子方法的发展,其中第一种方法最常用于常规诊断。日常分析核酸所依赖的方法依赖于扩增,以便能够特异性检测 SARS-CoV-2 RNA。本文旨在综述用于 SARS-CoV-2 检测的核酸扩增检测(NAATs)的最新进展,无论是在临床水平还是在即时护理(POC)水平,因此重点关注基于等温扩增和非等温扩增的诊断,同时仔细观察相关病毒学方面、检测步骤和仪器。在引言中进行主题背景介绍后,关于基础病毒学方面的基本知识、临床样本的采集和处理的主题为详细评估扩增和检测技术铺平了道路。为了讨论这些主题,讨论了核酸扩增方法、用于检测 DNA 的不同类型的分子反应以及执行此类分析途径所需的仪器。随后的部分讨论了具有代表性的商业检测的基准,这些基准建立在前几节中讨论的技术方面以及该部分提供的其他附加信息的基础上。最后几行旨在展望 NAAT 的未来及其在应对这场大流行和其他类似即将到来的挑战中的重要性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/adef/8984495/b2beb7e72a8d/fcimb-12-799678-g001.jpg

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