The coronavirus disease 2019 (COVID-19) has caused a large global outbreak. It is accordingly important to develop accurate and rapid diagnostic methods. The polymerase chain reaction (PCR)-based method including reverse transcription-polymerase chain reaction (RT-PCR) is the most widely used assay for the detection of SARS-CoV-2 RNA. Along with the RT-PCR method, digital PCR has emerged as a powerful tool to quantify nucleic acid of the virus with high accuracy and sensitivity. Non-PCR based techniques such as reverse transcription loop-mediated isothermal amplification (RT-LAMP) and reverse transcription recombinase polymerase amplification (RT-RPA) are considered to be rapid and simple nucleic acid detection methods and were reviewed in this paper. Non-conventional molecular diagnostic methods including next-generation sequencing (NGS), CRISPR-based assays and nanotechnology are improving the accuracy and sensitivity of COVID-19 diagnosis. In this review, we also focus on standardization of SARS-CoV-2 nucleic acid testing and the activity of the National Metrology Institutes (NMIs) and highlight resources such as reference materials (RM) that provide the values of specified properties. Finally, we summarize the useful resources for convenient COVID-19 molecular diagnostics.

中文翻译：

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SARS-CoV-2 核酸检测

2019 年冠状病毒病 (COVID-19) 已在全球范围内大规模爆发。因此，开发准确、快速的诊断方法非常重要。基于聚合酶链式反应 (PCR) 的方法，包括逆转录聚合酶链式反应 (RT-PCR)，是检测 SARS-CoV-2 RNA 时使用最广泛的检测方法。与 RT-PCR 方法一起，数字 PCR 已成为高精度和高灵敏度定量病毒核酸的有力工具。基于非PCR的技术，例如逆转录环介导的等温扩增（RT-LAMP）和逆转录重组酶聚合酶扩增（RT-RPA）被认为是快速且简单的核酸检测方法，本文对此进行了综述。包括下一代测序 (NGS)、基于 CRISPR 的检测和纳米技术在内的非常规分子诊断方法正在提高 COVID-19 诊断的准确性和灵敏度。在这篇综述中，我们还重点关注 SARS-CoV-2 核酸检测的标准化和国家计量研究所 (NMI) 的活动，并重点介绍提供特定属性值的参考材料 (RM) 等资源。最后，我们总结了方便的 COVID-19 分子诊断的有用资源。