RT-qPCR half-reaction optimization for the detection of SARS-CoV-2

The main laboratory test for the diagnosis of coronavirus disease 2019 (COVID-19) is the reverse transcription real-time polymerase chain reaction (RT-qPCR). However, RT-qPCR is expensive because of the number of tests required. This study aimed to evaluate an alternative to the RT-qPCR approach for...

Full description

Saved in:

Bibliographic Details
Published in:	Revista da Sociedade Brasileira de Medicina Tropical Vol. 54; p. e03192020
Main Authors:	Wink, Priscila Lamb, Volpato, Fabiana, Lima-Morales, Daiana de, Paiva, Rodrigo Minuto, Willig, Julia Biz, Bock, Hugo, Paris, Fernanda de, Barth, Afonso Luís
Format:	Journal Article
Language:	English
Published:	Brazil Sociedade Brasileira de Medicina Tropical - SBMT 01-01-2021 Sociedade Brasileira de Medicina Tropical (SBMT)
Subjects:	Coronavirus Coronavirus disease 2019 COVID-19 Humans Major Real-Time Polymerase Chain Reaction RNA, Viral - genetics RT-qPCR half-reaction SARS-CoV-2 Sensitivity and Specificity TROPICAL MEDICINE COVID-19 SARS-CoV-2 Coronavirus RT-qPCR half-reaction Coronavirus disease 2019
Online Access:	Get full text
Tags:	Add Tag No Tags, Be the first to tag this record!

Description
Summary:	The main laboratory test for the diagnosis of coronavirus disease 2019 (COVID-19) is the reverse transcription real-time polymerase chain reaction (RT-qPCR). However, RT-qPCR is expensive because of the number of tests required. This study aimed to evaluate an alternative to the RT-qPCR approach for the detection of sudden acute respiratory syndrome coronavirus 2 (SARS-CoV-2) that is half of the total volume currently recommended by the US Centers for Disease Control and Prevention. The analytical limit of detection (LoD) and the reaction efficiency using half volumes of the RT-qPCR assay were evaluated for the N1 and N2 regions using a synthetic control RNA. A panel of 76 SARS-CoV-2-positive and 26 SARS-CoV-2-negative clinical samples was evaluated to establish clinical sensitivity and specificity. The RT-qPCR assay efficiency was 105% for the half and standard reactions considering the N2 target and 84% (standard) and 101% (half) for N1. The RT-qPCR half-reaction LoD for N1 and N2 were 20 and 80 copies/µL, respectively. The clinical sensitivity and specificity were 100%. The half reaction presented a decrease of up to 5.5 cycle thresholds compared with standard RT-qPCR. The use of the RT-qPCR half-reaction proved feasible and economic for the detection of SARS-CoV-2 RNA.
Bibliography:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 Authors’ contribution: A.L.B is the principal investigator of this work and, as such, had full access to all of the data in the study and takes responsibility for the integrity of the data and the accuracy of the data analysis. Author's contribution: P.L.W. - Study conception and design, acquisition of data, drafting of manuscript, analysis and interpretation of data. F.V. - Analysis and interpretation of data. D.L.M. - Analysis and interpretation of data. R.M.P. - Study conception and design, acquisition of data, analysis and interpretation of data. J.B.W. - Study conception and design, acquisition of data, analysis and interpretation of data. H.B. - Analysis and interpretation of data. F.P. - Analysis and interpretation of data. A.L.B. - Critical revision. All authors critically reviewed. Conflict of Interest: The authors declare that there is no conflict of interest.
ISSN:	0037-8682 1678-9849 1678-9849
DOI:	10.1590/0037-8682-0319-2020