Diminished amplification of SARS-CoV-2 ORF1ab in a commercial dual-target qRT-PCR diagnostic assay

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Abstract

Here we describe a SARS-CoV-2 variant with diminished amplification of the ORF ORF1ab target in the Cobas® dual-target SARS-CoV-2 assay resulting in a discrepancy of Ct-values (Ct-value 20.7 for the E-gene and Ct-value 30.2 for ORF1ab). Five unique nucleotide mutations were identified in ORF1ab: C11450A (nsp10) C14178T (RdRp), G15006T (RdRp), G18394T (Hel), and G20995T (Hel). This case highlights the importance of surveillance of genomic regions used in molecular diagnostics and the importance of the public release of target regions used to update commercial and in-house developed SARS-CoV-2 PCR tests. This work underpins the importance of using dual-targets in molecular diagnostic assays to limit the change of false-negative results due to primer and/or probe mismatches.

Original languageEnglish
Article number114397
JournalJournal of Virological Methods
Volume300
DOIs
Publication statusPublished - Feb 2022

Bibliographical note

Funding Information:
SP: received financial support from Gilead Sciences outside of the submitted work.

Funding Information:
Sequencing was funded by ZonMW (grant agreement no: 10150062010005 ).

Publisher Copyright:
© 2021

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