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. 2020 May 27;6(5):591-605.
doi: 10.1021/acscentsci.0c00501. Epub 2020 Apr 30.

Assay Techniques and Test Development for COVID-19 Diagnosis

Linda J Carter  1 , Linda V Garner  1 , Jeffrey W Smoot  1 , Yingzhu Li  1 , Qiongqiong Zhou  1 , Catherine J Saveson  1 , Janet M Sasso  1 , Anne C Gregg  1 , Divya J Soares  1 , Tiffany R Beskid  1 , Susan R Jervey  1 , Cynthia Liu  1
Affiliations

Assay Techniques and Test Development for COVID-19 Diagnosis

Linda J Carter et al. ACS Cent Sci. .
No abstract available

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Figures

Figure 1
Figure 1
Reverse transcription-polymerase chain reaction (RT-PCR). The RT-PCR creates a cDNA copy of a specific segment of the viral RNA, which is converted to dsDNA that is exponentially amplified.
Figure 2
Figure 2
Reverse transcription loop-mediated isothermal amplification (RT-LAMP). Step 1: At the 3′-end of the viral RNA, reverse transcriptase and BIP primer initiate conversion of RNA to cDNA. Step 2: At the same end, DNA polymerase and B3 primer continue to generate the second cDNA strand to displace and release the first cDNA strand. Step 3: The FIP primer binds to the released cDNA strand and DNA polymerase generates the complementary strand. Step 4: F3 primer binds to the 3′ end, and DNA polymerase then generates a new strand while displacing the old strand. LAMP cycling produces various sized double-stranded looped DNA structures containing alternately inverted repeats of the target sequence as detected by a DNA indicator dye. Reagents*: Primers and master mix containing reverse transcriptase, DNA polymerase with strand displacing activity, dNTPs, and buffers.
Figure 3
Figure 3
Two alternative CRISPR methods for detecting viral RNA. Method A (SHERLOCK assay): RT-RPA (recombinase polymerase amplification) converts viral RNA to dsDNA. T7 transcription generates complementary RNA from the dsDNA template. The Cas13–tracrRNA complex binds to the target sequence, which activates the general nuclease enzyme activity of Cas13 to cleave the target sequence and the fluorescent RNA reporter. Method B (DETECTR assay): RT-RPA (recombinase polymerase amplification) converts viral RNA to dsDNA. The Cas12a–tracrRNA complex binds to the target sequence, which activates the general nuclease enzyme activity of Cas12a to cleave the target sequence and the fluorescent RNA reporter.
Figure 4
Figure 4
Nucleic acid hybridization using microarray. Viral cDNA and reference cDNA with different fluorescent labels are mixed and applied to the microarray wells coated with specific DNA probes.
Figure 5
Figure 5
ELISA assays detecting antibodies (A) or antigens (B).
Figure 6
Figure 6
Lateral flow immunoassay for detection of anti-SARS-CoV-2 antibodies. Samples move via capillary flow on the nitrocellulose membrane. When anti-SARS-CoV-2 antibodies are present, they bind to the labeled antigen and continue to move until they are captured by the immobilized antihuman antibodies. The presence of the captured antibody–antigen complex is visualized as a colored test band. The labeled control antibodies comigrate until they are captured at the control band.
Figure 7
Figure 7
Monthly trend of journal publications related to COVID-19 diagnostics in 2020.
See this image and copyright information in PMC

References

    1. Mizumoto K.; Kagaya K.; Zarebski A.; Chowell G. Estimating the asymptomatic proportion of coronavirus disease 2019 (COVID-19) cases on board the Diamond Princess cruise ship, Yokohama, Japan, 2020. Euro Surveill 2020, 25 (10), 2000180.10.2807/1560-7917.ES.2020.25.10.2000180. - DOI - PMC - PubMed
    1. Bendavid E.; Mulaney B.; Sood N.; Shah S.; Ling E.; Bromley-Dulfano R.; Lai C.; Weissberg Z.; Saavedra R.; Tedrow J.; Tversky D.; Bogan A.; Kupiec T.; Eichner D.; Gupta R.; Ioannidis J.; Bhattacharya J.. COVID-19 Antibody Seroprevalence in Santa Clara County, California medRxiv 2020, 2020.04.14.20062463.10.1101/2020.04.14.20062463 - DOI - PMC - PubMed
    1. Jones H.The Importance of diagnostic testing for COVID-19. Infectious Diseases Hub; April 2, 2020. www.id-hub.com/2020/04/02/the-importance-of-diagnostic-testing-for-covid....
    1. Tan R.COVID-19 Diagnostics Explained. Asian Scientist; April 8, 2020. www.asianscientist.com/2020/04/features/covid-19-diagnostics-explained/.
    1. Xia J.; Tong J.; Liu M.; Shen Y.; Guo D. Evaluation of coronavirus in tears and conjunctival secretions of patients with SARS-CoV-2 infection. J. Med. Virol. 2020, 92, 589–594. 10.1002/jmv.25725. - DOI - PMC - PubMed