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Brief comparative evaluation of six open one-step RT-qPCR mastermixes for the detection of SARS-CoV-2 RNA using a Taqman probe

Authors
  • Haddar, Cyrille1
  • Verhoeven, Paul O.2
  • Bourlet, Thomas2
  • Pozzetto, Bruno2
  • Pillet, Sylvie2
  • 1 BioSpeedia, Institut Pasteur, 75015, Paris, France
  • 2 Laboratory of Infectious Agents and Hygiene, University Hospital of Saint-Etienne, and GIMAP (Groupe Immunité des Muqueuses et Agents Pathogènes) EA-3064, Medicine Faculty of Saint-Etienne, Campus Santé-Innovations of Saint-Etienne, Member of University of Lyon, France
Type
Published Article
Journal
Journal of Clinical Virology
Publisher
Elsevier B.V.
Publication Date
Sep 08, 2020
Volume
132
Pages
104636–104636
Identifiers
DOI: 10.1016/j.jcv.2020.104636
PMID: 33099260
PMCID: PMC7476897
Source
PubMed Central
Keywords
License
Unknown

Abstract

Background Facing the emergence of a new RNA virus, clinical laboratories are often helpless in the case of a shortage of reagents recommended by Reference Centres. Objectives To compare five open one step RT-qPCR reagents to the SuperScript™ III Platinum™ One-Step qRT-PCR kit (Invitrogen) considered as the reference one in France at the beginning of the pandemic for detection of the Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) in respiratory specimens by using a laboratory-developed assay targeting the viral RNA dependant RNA polymerase (RdRp) gene. Study design A total of 51 NUCLISENS easyMAG extracts from respiratory specimens was tested on ABI 7500 thermocycler with TaqMan Fast Virus 1-Step Master Mix (Applied Biosystems), Luna® Universal Probe One-Step RT-qPCR Kit (New England Biolabs), GoTaq® Probe 1- Step RT-qPCR System (Promega), LightCycler® Multiplex RNA Virus Master (Roche) and One-step PrimeScript RT-PCR kit (Takara). The CT values obtained using the 5 challenged reagents were compared to those obtained using the reference assay. Results The percentages of concordance were all above 95 %. When comparing the CT values of the 48 extracts exhibiting CT values < 35 obtained with the reference reagent, the results were similar between the reagents although the differences of CT values were quite dispersed. Conclusions All five reagents can be considered as alternative reagents to the reference for detecting SARS-CoV-2 RNA.

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