SARS-CoV-2 Diagnostics Based on Nucleic Acids Amplification: From Fundamental Concepts to Applications and Beyond
TLDR
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.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.read more
Citations
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COVID-19 Diagnosis: A Comprehensive Review of the RT-qPCR Method for Detection of SARS-CoV-2
Debashis Dutta,Sarah Naiyer,Sabanaz Mansuri,Neeraj Soni,Vandana Singh,Khalid Hussain Bhat,Nishant Singh,Gunjan Arora,M. Shahid Mansuri +8 more
TL;DR: A comprehensive review of the natural history, pathology, molecular biology, and efficient diagnostic methods of COVID-19, detecting SARS-CoV-2 in collected samples and reviewing the gold standard RT-qPCR method to confer a better understanding and application to combat the CO VID-19 pandemic.
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The Future of Point-of-Care Nucleic Acid Amplification Diagnostics after COVID-19: Time to Walk the Walk
TL;DR: In this article , the main approaches to rapid and point-of-care diagnostics based on RT-qPCR and isothermal amplification diagnostics are discussed, with a particular focus on LAMP and recombinase polymerase amplification (RPA).
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Reverse transcriptase-free detection of viral RNA using Hemo Klentaq DNA polymerase
TL;DR: In this article , the authors used Hemo KlenTaq (HKTaq) as a DNA-dependent DNA polymerase, which possesses reverse transcriptase activity and provides reproducible amplification of RNA targets with an efficiency comparable to common reverse transcription (RT)PCR.
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A Molecular Lateral Flow Assay for SARS-CoV-2 Quantitative Detection
TL;DR: The developed test allowed for rapid detection by the naked eye and the simultaneous quantification of SARS-CoV-2 in nasopharyngeal swabs with high specificity, detectability, and repeatability.
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Low-Temperature Loop-Mediated Isothermal Amplification Operating at Physiological Temperature
Daehan Nam,Seokjoong Kim,Jung Ho Kim,Seungjin Lee,Jinseo Son,Doyeon Kim,Byung Seok Cha,Eun Sung Lee,Ki Soo Park +8 more
TL;DR: Low-temperature LAMP as discussed by the authors operates at a physiological temperature of 37 °C, which is similar to that of conventional LAMP and achieves a similar amplification efficiency as that of LAMP at 65 °C.
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