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SARS-CoV-2 RNA detection using pooling of self-collected samples: Simple protocols may foster asymptomatic surveillance

Lopes, G. I. S. L.; Carmona, R. d. C. C.; Silva, V. O.; Ahagon, C. M.; do Prado, L. S.; dos Santos, F. C. P.; da Silva, D. B. B.; Ribeiro, W. N.; Matshuda, E. M.; Cilli, A.; Afonso, A. M. S.; Pinho, M. A. B.; Timnetsky, M. d. C. S. T.; Brigido, L. F. d. M.

2020-10-07 infectious diseases
10.1101/2020.10.05.20205872 medRxiv
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1BackgroundSurveillance of COVID infection and isolation of infected individuals is one of the available tools to control the spread of SAR-CoV-2. Asymptomatic and pre symptomatic are responsible for substantial transmission. RNA or antigen tests are necessary to identify non-symptomatic individuals. We tested the feasibility of using samples pooling offering different collection alternatives (swab/throat wash/saliva) to volunteers of a public health institute. MethodsWe evaluated pool samples from frozen material from previously tested samples and a prospective collection from asymptomatic volunteers. Some collections were paired for comparison. Pools and some individual samples were extracted with QIAamp Viral RNA Mini Kit (Qiagen, USA) and/or Lucigen Quick Extract DNA extraction solution (BioSearch, USA) and submitted to rtPCR (Allplex, Seegene, Korea). ResultsA total of 240 samples from 130 new collections and 37 samples with known result were evaluated. Pool CT was generally higher than individual samples. Lucigen extraction showed higher CT, including false negative results for samples with high CT at Qiagen extraction. Paired Swab and TW samples showed comparable results. No volunteer from negative pools reported any symptom in the 2-3 days after collection. ConclusionsClinical samples pooling to detect SARS-CoV-2 RNA is feasible and an economical way to test for COVID-19, especially in surveillance strategies targeting more infectiousness, higher viremia individuals. The use of Lucigen reagents show lower sensibility that may lead to false negative results with lower viremia samples. Combining throat wash with saliva may provide and interesting self-collection alternative, but more comparative work is needed.

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