RT info:eu-repo/semantics/article
T1 Increasing SARS-CoV-2 RT-qPCR testing capacity by sample pooling.
A1 Valenzuela Fernández, Agustín
A1 Alcoba Florez, Julia
A1 Gil Campesino, Helena
A1 García Martínez de Artola, Diego
A1 Díez Gil, Óscar
A1 González Montelongo, Rafaela
A1 Ciuffreda, Laura
A1 Flores, Carlos
A2 Medicina Física y Farmacología
A2 Grupo "inmunología Celular y Viral".
K1 COVID-19
K1 SARS-CoV-2
K1 testing capacity
K1 scalability
K1 sample pooling
AB Objectives: Limited testing capacity has characterized the ongoing COVID-19pandemic in Spain, hampering a timely control of outbreaks and the possibilities toreduce the escalation of community transmissions. Here we investigated the potential ofusing pooling of samples followed by one-step retrotranscription and quantitative PCR(RT-qPCR) to increase SARS-CoV-2 testing capacity.Methods: We first evaluated different sample pooling (1:5, 1:10 and 1:15) prior to RNAextractions followed by standard RT-qPCR for SARS-CoV-2/COVID-19 diagnosis.The pool size achieving reproducible results in independent tests was then used forassessing nasopharyngeal samples in a tertiary hospital during August 2020.Results: We found that pool size of five samples achieved the highest sensitivitycompared to pool sizes of 10 and 15, showing a mean (± SD) Ct shift of 3.5 ± 2.2between the pooled test and positive samples in the pool. We then used a pool size offive to test a total of 895 pools (4,475 prospective samples) using two different RTqPCR kits available at that time. The Real Accurate Quadruplex corona-plus PCR Kit(PathoFinder) reported the lowest mean Ct (± SD) shift (2.2 ± 2.4) among the pool andthe individual samples. The strategy allows detecting individual samples in the positivepools with Cts in the range of 16.7-39.4.Conclusions: We found that pools of five samples combined with RT-qPCR solutionshelped to increase SARS-CoV-2 testing capacity with minimal loss of sensitivitycompared to that resulting from testing the samples independently.
YR 2020
FD 2020
LK http://riull.ull.es/xmlui/handle/915/35347
UL http://riull.ull.es/xmlui/handle/915/35347
LA en
NO medRxiv preprint doi: https://doi.org/10.1101/2020.10.24.20218685; this version posted October 27, 2020. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity.. It is made available under a CC-BY-NC-ND 4.0 International license .
DS Repositorio institucional de la Universidad de La Laguna
RD 24-nov-2024