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A Novel Saliva RT-LAMP Workflow for Rapid Identification of COVID-19 Cases and Restraining Viral Spread

1
Centro de Pesquisa Sobre o Genoma Humano e Células-Tronco (HUG-CELL), Instituto de Biociências, Universidade de São Paulo (USP), São Paulo 05508-090, Brazil
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Instituto de Medicina Tropical, Universidade de São Paulo (USP), São Paulo 05403-000, Brazil
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Grupo Fleury, Research and Development, São Paulo 04344-070, Brazil
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Instituto de Ensino e Pesquisa Prevent Senior, São Paulo 04547-100, Brazil
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Faculdade de Medicina, Universidade Municipal de São Caetano do Sul (USCS), São Paulo 09521-160, Brazil
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Instituto de Química, Universidade de São Paulo (USP), São Paulo 05508-000, Brazil
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Faculdade de Ciências Farmacêuticas de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto 14040-903, Brazil
*
Authors to whom correspondence should be addressed.
Diagnostics 2021, 11(8), 1400; https://doi.org/10.3390/diagnostics11081400
Received: 8 June 2021 / Revised: 25 June 2021 / Accepted: 16 July 2021 / Published: 3 August 2021
(This article belongs to the Section Diagnostic Microbiology and Infectious Disease)
Rapid diagnostics is pivotal to curb SARS-CoV-2 transmission, and saliva has emerged as a practical alternative to naso/oropharyngeal (NOP) specimens. We aimed to develop a direct RT-LAMP (reverse transcription loop-mediated isothermal amplification) workflow for viral detection in saliva, and to provide more information regarding its potential in curbing COVID-19 transmission. Clinical and contrived specimens were used to optimize formulations and sample processing protocols. Salivary viral load was determined in symptomatic patients to evaluate the clinical performance of the test and to characterize saliva based on age, gender and time from onset of symptoms. Our workflow achieved an overall sensitivity of 77.2% (n = 90), with 93.2% sensitivity, 97% specificity, and 0.895 Kappa for specimens containing >102 copies/μL (n = 77). Further analyses in saliva showed that viral load peaks in the first days of symptoms and decreases afterwards, and that viral load is ~10 times lower in females compared to males, and declines following symptom onset. NOP RT-PCR data did not yield relevant associations. This work suggests that saliva reflects the transmission dynamics better than NOP specimens, and reveals gender differences that may reflect higher transmission by males. This saliva RT-LAMP workflow can be applied to track viral spread and, to maximize detection, testing should be performed immediately after symptoms are presented, especially in females. View Full-Text
Keywords: LAMP (loop-mediated isothermal amplification) assay; 2019 novel coronavirus; saliva; viral diagnostics LAMP (loop-mediated isothermal amplification) assay; 2019 novel coronavirus; saliva; viral diagnostics
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MDPI and ACS Style

Kobayashi, G.S.; Brito, L.A.; Moreira, D.d.P.; Suzuki, A.M.; Hsia, G.S.P.; Pimentel, L.F.; de Paiva, A.P.B.; Dias, C.R.; Lourenço, N.C.V.; Oliveira, B.A.; Manuli, E.R.; Corral, M.A.; Cavaçana, N.; Mitne-Neto, M.; Sales, M.M.; Dell’ Aquila, L.P.; Filho, A.R.; Parrillo, E.F.; Mendes-Corrêa, M.C.; Sabino, E.C.; Costa, S.F.; Leal, F.E.; Sgro, G.G.; Farah, C.S.; Zatz, M.; Passos-Bueno, M.R. A Novel Saliva RT-LAMP Workflow for Rapid Identification of COVID-19 Cases and Restraining Viral Spread. Diagnostics 2021, 11, 1400. https://doi.org/10.3390/diagnostics11081400

AMA Style

Kobayashi GS, Brito LA, Moreira DdP, Suzuki AM, Hsia GSP, Pimentel LF, de Paiva APB, Dias CR, Lourenço NCV, Oliveira BA, Manuli ER, Corral MA, Cavaçana N, Mitne-Neto M, Sales MM, Dell’ Aquila LP, Filho AR, Parrillo EF, Mendes-Corrêa MC, Sabino EC, Costa SF, Leal FE, Sgro GG, Farah CS, Zatz M, Passos-Bueno MR. A Novel Saliva RT-LAMP Workflow for Rapid Identification of COVID-19 Cases and Restraining Viral Spread. Diagnostics. 2021; 11(8):1400. https://doi.org/10.3390/diagnostics11081400

Chicago/Turabian Style

Kobayashi, Gerson S., Luciano A. Brito, Danielle d.P. Moreira, Angela M. Suzuki, Gabriella S.P. Hsia, Lylyan F. Pimentel, Ana P.B. de Paiva, Carolina R. Dias, Naila C.V. Lourenço, Beatriz A. Oliveira, Erika R. Manuli, Marcelo A. Corral, Natale Cavaçana, Miguel Mitne-Neto, Maria M. Sales, Luiz P. Dell’ Aquila, Alvaro R. Filho, Eduardo F. Parrillo, Maria C. Mendes-Corrêa, Ester C. Sabino, Silvia F. Costa, Fabio E. Leal, Germán G. Sgro, Chuck S. Farah, Mayana Zatz, and Maria R. Passos-Bueno 2021. "A Novel Saliva RT-LAMP Workflow for Rapid Identification of COVID-19 Cases and Restraining Viral Spread" Diagnostics 11, no. 8: 1400. https://doi.org/10.3390/diagnostics11081400

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