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, H.
A1 García-Martínez de Artola, D.
A1 Díez-Gil, O.
A1 González-Montelongo, R.
A1 Ciuffreda, L.
A1 Flores, C.
K1 COVID-19
K1 SARS-CoV-2
K1 Testing capacity
K1 Scalability
K1 Sample pooling
AB Objectives: Limited testing capacity has characterized the ongoing coronavirus disease 2019 (COVID-19) pandemic in Spain, hampering timely control of outbreaks and opportunities to reduce the escalation of community transmission. This study investigated the potential to use sample pooling, followed by one-step retrotranscription and real-time quantitative reverse transcription polymerase chain reaction (RT-qPCR) to increase testing capacity for severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2). Methods: Various pool sizes (five, 10 and 15 samples) were evaluated prior to RNA extraction followed by standard RT-qPCR for the diagnosis of COVID-19. The pool size achieving reproducible results with individual sample testing was subsequently used to assess nasopharyngeal samples in a tertiary hospital in August 2020. Results: A pool size of five samples had higher sensitivity compared with pool sizes of 10 and 15 samples, showing a mean cycle threshold (Ct) shift of 3.5 [standard deviation (SD) 2.2] between the pooled test and positive samples in the pool. Next, a pool size of five was used to test a total of 895 pools (4475 prospective samples) using two different RT-qPCR kits. The Real Accurate Quadruplex corona-plus PCR Kit (PathoFinder) reported the lowest mean Ct shift [2.2 (SD 2.4)] between the pool and individual samples. This strategy enables detection of individual positive samples in positive pools with Ct of 16.7–39.4. Conclusions: Grouping samples into pools of five for RT-qPCR resulted in an increase in SARS-CoV-2 testing capacity with minimal loss of sensitivity compared with testing each sample individually.
SN 1878-3511
YR 2021
FD 2021
LK http://riull.ull.es/xmlui/handle/915/35344
UL http://riull.ull.es/xmlui/handle/915/35344
LA en
DS Repositorio institucional de la Universidad de La Laguna
RD 20-may-2024