Comparison of eleven in vitro diagnostic assays for the detection of SARS-CoV-2 RNA.

Autor: Dundon WG; Animal Production and Health Laboratory, Joint FAO/IAEA Centre for Nuclear Applications in Food and Agriculture, Department of Nuclear Sciences and Applications, International Atomic Energy Agency, Friedenstrasse 1, A-2444, Seibersdorf, Austria. Electronic address: w.dundon@iaea.org., Settypalli TBK; Animal Production and Health Laboratory, Joint FAO/IAEA Centre for Nuclear Applications in Food and Agriculture, Department of Nuclear Sciences and Applications, International Atomic Energy Agency, Friedenstrasse 1, A-2444, Seibersdorf, Austria., Spiegel K; Department for Molecular Biology, Institute for Veterinary Disease Control, Austrian Agency for Health and Food Safety (AGES), Robert Koch Gasse 17, A-2340, Mödling, Austria., Steinrigl A; Department for Molecular Biology, Institute for Veterinary Disease Control, Austrian Agency for Health and Food Safety (AGES), Robert Koch Gasse 17, A-2340, Mödling, Austria., Revilla-Fernández S; Department for Molecular Biology, Institute for Veterinary Disease Control, Austrian Agency for Health and Food Safety (AGES), Robert Koch Gasse 17, A-2340, Mödling, Austria., Schmoll F; Department for Molecular Biology, Institute for Veterinary Disease Control, Austrian Agency for Health and Food Safety (AGES), Robert Koch Gasse 17, A-2340, Mödling, Austria., Naletoski I; Animal Production and Health Laboratory, Joint FAO/IAEA Centre for Nuclear Applications in Food and Agriculture, Department of Nuclear Sciences and Applications, International Atomic Energy Agency, Friedenstrasse 1, A-2444, Seibersdorf, Austria., Lamien CE; Animal Production and Health Laboratory, Joint FAO/IAEA Centre for Nuclear Applications in Food and Agriculture, Department of Nuclear Sciences and Applications, International Atomic Energy Agency, Friedenstrasse 1, A-2444, Seibersdorf, Austria., Cattoli G; Animal Production and Health Laboratory, Joint FAO/IAEA Centre for Nuclear Applications in Food and Agriculture, Department of Nuclear Sciences and Applications, International Atomic Energy Agency, Friedenstrasse 1, A-2444, Seibersdorf, Austria.
Jazyk: angličtina
Zdroj: Journal of virological methods [J Virol Methods] 2021 Sep; Vol. 295, pp. 114200. Date of Electronic Publication: 2021 Jun 01.
DOI: 10.1016/j.jviromet.2021.114200
Abstrakt: Transmission mitigation of SARS-CoV-2 requires the availability of accurate and sensitive detection methods. There are several commercial ad hoc molecular diagnostic kits currently on the market, many of which have been evaluated by different groups. However, in low resource settings the availability and cost of these commercial kits can be a limiting factor for many diagnostic laboratories. In such cases alternatives need to be identified. With this in mind, eight commercial reverse transcription quantitative real-time PCR (RT-qPCR) master mixes from Applied Biosystems (Thermo Fisher Scientific), Bio-Rad, Biotech Rabbit, Promega, Qiagen, QuantaBio, Invitrogen (Thermo Fisher Scientific) and Takara using the same commercial primer and probe mix [LightMix® Modular SARS and Wuhan CoV E-gene mix (TIB MolBiol, Germany)] were evaluated. Three ad hoc molecular diagnostic kits [GeneFinder™ COVID-19 Plus RealAmp kit (Osang Healthcare); genesig® Real-Time PCR Coronavirus COVID-19 (Primerdesign); and ViroReal® Kit SARS-CoV-2 & SARS-CoV (Ingenetix)] were also included in the study. The limit of detection was calculated for each assay using serial dilutions of a defined clinical sample. The performances of the assays were compared using a panel of 178 clinical samples and their analytical specificity assessed against a panel of human betacoronaviruses. Inter assay agreement was assessed using statistical tests (Bland-Altman, Fleiss-Kappa and Cohen's Kappa) and was shown to be excellent to good in all cases. We conclude that all of the assays evaluated in this study can be used for the routine detection of SARS-CoV-2 and that the RT-qPCR master mixes are a valid alternative to ad hoc molecular diagnostic kits.
(Copyright © 2021 The Author(s). Published by Elsevier B.V. All rights reserved.)
Databáze: MEDLINE
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