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. 2022 Jun;2(2):100074.
doi: 10.1016/j.jcvp.2022.100074. Epub 2022 Mar 24.

Evaluation of a novel direct RT-LAMP assay for the detection of SARS-CoV-2 from saliva samples in asymptomatic individuals

Benjamin Brown  1 , Robert William O'Hara  1 , Malcolm Guiver  1 , Emma Davies  1 , Andrew Birtles  2 , Hamzah Farooq  2 , Arpana Verma  3 , Hui Guo  3 , Katharine Hayden  2 , Nicholas Machin  1   2   3
Affiliations

Evaluation of a novel direct RT-LAMP assay for the detection of SARS-CoV-2 from saliva samples in asymptomatic individuals

Benjamin Brown et al. J Clin Virol Plus. 2022 Jun.

Abstract

Large scale screening of health care workers and the general population for asymptomatic COVID-19 infection requires modalities that are amenable to testing at scale while retaining acceptable levels of sensitivity and specificity. This study evaluated a novel COVID-19 Direct-RT LAMP assay using saliva samples in asymptomatic individuals by comparison to RT-PCR. Additional studies were performed using VTM collected from routine diagnostic testing. Analytical sensitivity was determined for Direct RT-LAMP assay using the WHO International Standard. Finally, quantified results from RT-PCR testing of 9177 nose and throat swabs obtained from routine diagnostic testing were used to estimate the sensitivity of Direct RT-LAMP using the limit of detection curve obtained from the analytical sensitivity data. Results from saliva testing demonstrated a sensitivity of 40.91% and a specificity of 100% for Direct RT-LAMP. The sensitivity and specificity for nose and throat swabs were 44.85% and 100% respectively. The 95% limit of detection (LOD) for Direct RT-LAMP was log 7.13 IU/ml (95% 6.9-7.5). The estimated sensitivity for Direct-RT LAMP based on the results of 9117 nose and throat swabs was 34% and 45% for saliva and VTM respectively. The overall diagnostic sensitivity of Direct RT-LAMP was low compared to RT-PCR. Testing of nose and throat swabs and estimating the sensitivity based on a large cohort of clinical samples demonstrated similar results. This study highlights the importance of utilising the prospective collection of samples from the intended target population in the assessment of diagnostic sensitivity.

Keywords: Asymptomaitc; COVID-19; Direct RT-LAMP; Real-time PCR; SARS-CoV-2; Saliva; Screening.

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Conflict of interest statement

The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

Figures

Fig. 1
Fig. 1
Dot distribution plot of the N1N2 assay Ct. values from confirmed RT-PCR positive saliva samples. Red circles were positive by RT-PCR and Direct RT-LAMP, clear circles are RT-PCR positive only.
Fig. 2
Fig. 2
Dot distribution plot of the N1N2 assay Ct. values from confirmed RT-PCR positive nose and throat swabs. Red circles represent specimens positive by both RT-PCR and Direct RT-LAMP, clear circles positive by RT-PCR only.
Fig 3
Fig. 3
Distribution of viral loads from RT-PCR positive samples and% LOD curves for Direct RT-LAMP testing of saliva and VTM.
See this image and copyright information in PMC

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References

    1. Hoehl S., et al. Evidence of SARS-CoV-2 infection in returning travelers from Wuhan, China. N. Engl. J. Med. 2020;382:1278. - PMC - PubMed
    1. Rivett L., et al. Screening of healthcare workers for SARS-CoV-2 highlights the role of asymptomatic carriage in COVID-19 transmission. Elife. 2020;9 - PMC - PubMed
    1. Arons M.M., et al. Presymptomatic SARS-CoV-2 infections and transmission in a skilled nursing facility. N. Engl. J. Med. 2020;382:2081–2090. - PMC - PubMed
    1. Ellingford J.M., et al. Genomic and healthcare dynamics of nosocomial sars-cov-2 transmission. Elife. 2021;10 - PMC - PubMed
    1. NICE . COVID-19 Rapid guideline: Haematopoietic Stem Cell Transplantation. 2020. Overview | COVID-19 rapid guideline: haematopoietic stem cell transplantation | Guidance | NICE.