Point-of-Care PCR Assays for COVID-19 Detection.

Niharika Gupta,Tarun Narayan,John H T Luong,Alan O’Riordan,D Kumar,Shine Augustine,Bansi D Malhotra,Asmita Das

doi:10.3390/bios11050141

Abstract

Molecular diagnostics has been the front runner in the world’s response to the COVID-19 pandemic. Particularly, reverse transcriptase-polymerase chain reaction (RT-PCR) and the quantitative variant (qRT-PCR) have been the gold standard for COVID-19 diagnosis. However, faster antigen tests and other point-of-care (POC) devices have also played a significant role in containing the spread of SARS-CoV-2 by facilitating mass screening and delivering results in less time. Thus, despite the higher sensitivity and specificity of the RT-PCR assays, the impact of POC tests cannot be ignored. As a consequence, there has been an increased interest in the development of miniaturized, high-throughput, and automated PCR systems, many of which can be used at point-of-care. This review summarizes the recent advances in the development of miniaturized PCR systems with an emphasis on COVID-19 detection. The distinct features of digital PCR and electrochemical PCR are detailed along with the challenges. The potential of CRISPR/Cas technology for POC diagnostics is also highlighted. Commercial RT–PCR POC systems approved by various agencies for COVID-19 detection are discussed.

Highlights

The coronavirus disease 2019 (COVID-19) outbreak crisis has changed the shape of our world since its first report in December 2019
reverse transcriptase-polymerase chain reaction (RT-PCR) is the first molecular diagnostic test to be employed for detecting SARS-CoV-2 RNA in patient samples and is currently considered the gold standard for COVID-19 diagnosis
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Summary

Introduction

The coronavirus disease 2019 (COVID-19) outbreak crisis has changed the shape of our world since its first report in December 2019. Faster, miniaturized isothermal amplification tests emerged that could detect the virus within a few minutes and with sensitivity at par with RT-PCR assays [5,9,10]. Different types of POCT devices have been authorized in various countries for emergency use, many novel biosensing strategies and designs still seek validation and are currently subject to academic inquiry. These devices have shorter response times and have cost-effectively enabled populationwide mass screening. The high temperature requirements of RT-PCR prevent non-specific amplification, which is more common in isothermal amplification techniques These temperature requirements somewhat complicate the development of PCR-based rapid devices. The potential of biosensing formats to be integrated with RT-PCR is explored, along with the path-breaking integration of CRISPR/Cas technology with amplification assays toward the development of faster, miniaturized devices and chips

RT-PCR

Electrochemical PCR

Future Outlook

Findings

Conclusions