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Article

Adjusting the Cut-Off and Maximum Pool Size in RT-qPCR Pool Testing for SARS-CoV-2

1
Graduate Program in Infectology and Tropical Medicine, Medical School, Universidade Federal de Minas Gerais, Belo Horizonte 30130-100, MG, Brazil
2
Vaccine Technology Center, Universidade Federal de Minas Gerais, Belo Horizonte 31310-260, MG, Brazil
3
Graduate Program in Health and Nutrition, Universidade Federal de Ouro Preto, Ouro Preto 35400-000, MG, Brazil
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Department of Preventive and Social Medicine, Medical School, Universidade Federal de Minas Gerais, Belo Horizonte 30130-100, MG, Brazil
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Department of Pediatrics, Medical School, Universidade Federal de Minas Gerais, Belo Horizonte 30130-100, MG, Brazil
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Department of Mathematics, Universidade Federal de Minas Gerais, Belo Horizonte 31270-901, MG, Brazil
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Department of Internal Medicine, Medical School, Universidade Federal de Minas Gerais, Belo Horizonte 30130-100, MG, Brazil
*
Author to whom correspondence should be addressed.
Academic Editor: Stefano Aquaro
Viruses 2021, 13(4), 557; https://doi.org/10.3390/v13040557
Received: 5 February 2021 / Revised: 15 March 2021 / Accepted: 23 March 2021 / Published: 26 March 2021
(This article belongs to the Special Issue Viral Infections in Developing Countries)
Reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR) to detect SARS-CoV-2 RNA is an essential test to monitor the occurrence of COVID-19. A methodology is proposed for the determination of maximum pool size and adjustments of cut-off values of cycle threshold (Ct in RT-qPCR pool testing, to compensate for the dilution caused by pooling. The trade-off between pool size and test sensitivity is stated explicitly. The procedure was designed to ensure that samples that would be detectable in individual testing remain detectable in pool testing. The proposed relaxation in cut-off is dependent on the pool size, allowing a relatively tight correction to avoid loss of detection of positive samples. The methodology was evaluated in a study of pool testing of adults attending a public emergency care unit, reference for COVID-19 in Belo Horizonte, Brazil, and presenting flu-like symptoms. Even samples on the edge of detectability in individual testing were detected correctly. The proposed procedure enhances the consistency of RT-qPCR pool testing by enforcing that the scales of detectability in pool processing and in individual sample processing are compatible. This may enhance the contribution of pool testing to large-scale testing for COVID-19. View Full-Text
Keywords: SARS-CoV-2; COVID-19; pool testing; RT-qPCR SARS-CoV-2; COVID-19; pool testing; RT-qPCR
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MDPI and ACS Style

Costa, M.S.; Sato, H.I.; Rocha, R.P.; Carvalho, A.F.; Guimarães, N.S.; Machado, E.L.; Alves, C.R.L.; Teixeira, S.M.R.; Takahashi, R.H.C.; Tupinambás, U.; da Fonseca, F.G. Adjusting the Cut-Off and Maximum Pool Size in RT-qPCR Pool Testing for SARS-CoV-2. Viruses 2021, 13, 557. https://doi.org/10.3390/v13040557

AMA Style

Costa MS, Sato HI, Rocha RP, Carvalho AF, Guimarães NS, Machado EL, Alves CRL, Teixeira SMR, Takahashi RHC, Tupinambás U, da Fonseca FG. Adjusting the Cut-Off and Maximum Pool Size in RT-qPCR Pool Testing for SARS-CoV-2. Viruses. 2021; 13(4):557. https://doi.org/10.3390/v13040557

Chicago/Turabian Style

Costa, Murilo S., Hugo I. Sato, Raissa P. Rocha, Alex F. Carvalho, Nathalia S. Guimarães, Elaine L. Machado, Claudia R.L. Alves, Santuza M.R. Teixeira, Ricardo H.C. Takahashi, Unaí Tupinambás, and Flávio G. da Fonseca. 2021. "Adjusting the Cut-Off and Maximum Pool Size in RT-qPCR Pool Testing for SARS-CoV-2" Viruses 13, no. 4: 557. https://doi.org/10.3390/v13040557

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