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Review
. 2021 May 1;11(5):141.
doi: 10.3390/bios11050141.

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

Niharika Gupta  1 , Shine Augustine  1 , Tarun Narayan  2 , Alan O'Riordan  2 , Asmita Das  1 , D Kumar  3 , John H T Luong  4 , Bansi D Malhotra  1
Affiliations
Review

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

Niharika Gupta et al. Biosensors (Basel). .

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.

Keywords: COVID-19; digital PCR; electrochemical; point-of-care; polymerase chain reaction.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Schematic diagram depicting the various steps performed by the CovidNudge assay for automated detection of SARS-CoV-2 RNA (Reprinted with permission from Ref. [20]).
Figure 2
Figure 2
Schematic depicting workflow of a ddPCR system: (A) preparation for amplification, (B) generation of water-in-oil droplets using a microfluidic flow system, (C) collection of the droplets in PCR tubes, (D) PCR amplification, (E) analysis of fluorescence in the droplets after amplification, and (F) fitting to Poisson distribution to determine the absolute copy numbers of the target molecules (Reprinted from Ref. [28]).
Figure 3
Figure 3
Workflow of the RCA-based electrochemical sensor for SARS-CoV-2 detection (Reprinted from Ref. [50]).
See this image and copyright information in PMC

References

    1. COVID-19 Weekly Epidemiological Update, 9 March 2021. World Health Organization; Geneva, Switzerland: 2021.
    1. Arnaout R., Lee R.A., Lee G.R., Callahan C., Yen C.F., Smith K.P., Arora R., Kirby J.E. SARS-CoV2 testing: The limit of detection matters. bioRxiv. 2020 doi: 10.1101/2020.06.02.131144. - DOI - PMC - PubMed
    1. Grant B.D., Anderson C.E., Williford J.R., Alonzo L.F., Glukhova V.A., Boyle D.S., Weigl B.H., Nichols K.P. SARS-CoV-2 coronavirus nucleocapsid antigen-detecting half-strip lateral flow assay toward the development of point of care tests using commercially available reagents. Anal. Chem. 2020;92:11305–11309. doi: 10.1021/acs.analchem.0c01975. - DOI - PubMed
    1. Ragnesola B., Jin D., Lamb C.C., Shaz B.H., Hillyer C.D., Luchsinger L.L. COVID19 antibody detection using lateral flow assay tests in a cohort of convalescent plasma donors. BMC Res. Notes. 2020;13:1–7. doi: 10.1186/s13104-020-05212-0. - DOI - PMC - PubMed
    1. Zhu X., Wang X., Han L., Chen T., Wang L., Li H., Li S., He L., Fu X., Chen S. Multiplex reverse transcription loop-mediated isothermal amplification combined with nanoparticle-based lateral flow biosensor for the diagnosis of COVID-19. Biosens. Bioelectron. 2020;166:112437. doi: 10.1016/j.bios.2020.112437. - DOI - PMC - PubMed

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