Comparison of SARS-CoV2 N gene real-time RT-PCR targets and commercially available mastermixes

Author:

Brown Julianne RORCID,O’Sullivan Denise,Pereira Rui PA,Whale Alexandra S,Busby Eloise,Huggett Jim,Harris Kathryn

Abstract

ABSTRACTWe aim to test four one-step RT real-time mastermix options for use in SARS-CoV2 real-time PCR, with three primer/probe assays targeting the N gene. The lower limit of detection is determined using a SARS CoV2 N gene RNA transcript dilution series (to 1 copy/µl) and verified using 74 nose and throat swabs.The N2 assay demonstrates the most sensitive detection of SARS-Cov-2 RNA. Three of the four mastermixes performed well, with the Takara One Step PrimeScript™ III RT-PCR Kit mastermix demonstrating improved performance at the lower limit of detection.

Publisher

Cold Spring Harbor Laboratory

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