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Deprecated: Implicit conversion from float 251.2 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 J+Clin+Virol 2020 ; 132 (ä): 104636 Nephropedia Template TP
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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 #MMPMID33099260
Haddar C; Verhoeven PO; Bourlet T; Pozzetto B; Pillet S
J Clin Virol 2020[Nov]; 132 (ä): 104636 PMID33099260show ga
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(R) Universal Probe One-Step RT-qPCR Kit (New England Biolabs), GoTaq(R) Probe 1- Step RT-qPCR System (Promega), LightCycler(R) 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.