tetano
Editor, Senior Moderator
Sci Rep
. 2022 Jun 27;12(1):10867.
doi: 10.1038/s41598-022-13995-4.
Emergence of a mutation in the nucleocapsid gene of SARS-CoV-2 interferes with PCR detection in Canada
Sandra Isabel[SUP] 1 [/SUP], Mariana Abdulnoor[SUP] 1 2 [/SUP], Karel Boissinot[SUP] 3 [/SUP], Marc R Isabel[SUP] 1 4 [/SUP], Richard de Borja[SUP] 5 [/SUP], Philip C Zuzarte[SUP] 5 [/SUP], Calvin P Sjaarda[SUP] 6 7 [/SUP], Kevin R Barker[SUP] 1 8 9 [/SUP], Prameet M Sheth[SUP] 10 11 12 [/SUP], Larissa M Matukas[SUP] 1 3 [/SUP], Jonathan B Gubbay[SUP] 1 2 13 [/SUP], Alisson J McGeer[SUP] 1 14 [/SUP], Samira Mubareka[SUP] 1 15 [/SUP], Jared T Simpson[SUP] 5 16 [/SUP], Ramzi Fattouh[SUP] 17 18 19 [/SUP]
Affiliations
Abstract
The emergence of Severe Acute Respiratory Syndrome Coronavirus (SARS-CoV-2) was met with rapid development of robust molecular-based detection assays. Many SARS-CoV-2 molecular tests target multiple genetic regions of the virus to maximize detection and protect against diagnostic escape. Despite the relatively moderate mutational rate of SARS-CoV-2, numerous mutations with known negative impact on diagnostic assays have been identified. In early 2021, we identified four samples positive for SARS-CoV-2 with a nucleocapsid (N) gene drop out on Cepheid Xpert® Xpress SARS-CoV-2 assay. Sequencing revealed a single common mutation in the N gene C29200T. Spatiotemporal analysis showed that the mutation was found in at least six different Canadian provinces from May 2020 until May 2021. Phylogenetic analysis showed that this mutation arose multiple times in Canadian samples and is present in six different variants of interest and of concern. The Cepheid testing platform is commonly used in Canada including in remote regions. As such, the existence of N gene mutation dropouts required further investigation. While commercial SARS-CoV-2 molecular detection assays have contributed immensely to the response effort, many vendors are reluctant to make primer/probe sequences publicly available. Proprietary primer/probe sequences create diagnostic 'blind spots' for global SARS-CoV-2 sequence monitoring and limits the ability to detect and track the presence and prevalence of diagnostic escape mutations. We hope that our industry partners will seriously consider making primer/probe sequences available, so that diagnostic escape mutants can be identified promptly and responded to appropriately to maintain diagnostic accuracy.
. 2022 Jun 27;12(1):10867.
doi: 10.1038/s41598-022-13995-4.
Emergence of a mutation in the nucleocapsid gene of SARS-CoV-2 interferes with PCR detection in Canada
Sandra Isabel[SUP] 1 [/SUP], Mariana Abdulnoor[SUP] 1 2 [/SUP], Karel Boissinot[SUP] 3 [/SUP], Marc R Isabel[SUP] 1 4 [/SUP], Richard de Borja[SUP] 5 [/SUP], Philip C Zuzarte[SUP] 5 [/SUP], Calvin P Sjaarda[SUP] 6 7 [/SUP], Kevin R Barker[SUP] 1 8 9 [/SUP], Prameet M Sheth[SUP] 10 11 12 [/SUP], Larissa M Matukas[SUP] 1 3 [/SUP], Jonathan B Gubbay[SUP] 1 2 13 [/SUP], Alisson J McGeer[SUP] 1 14 [/SUP], Samira Mubareka[SUP] 1 15 [/SUP], Jared T Simpson[SUP] 5 16 [/SUP], Ramzi Fattouh[SUP] 17 18 19 [/SUP]
Affiliations
- PMID: 35760824
- DOI: 10.1038/s41598-022-13995-4
Abstract
The emergence of Severe Acute Respiratory Syndrome Coronavirus (SARS-CoV-2) was met with rapid development of robust molecular-based detection assays. Many SARS-CoV-2 molecular tests target multiple genetic regions of the virus to maximize detection and protect against diagnostic escape. Despite the relatively moderate mutational rate of SARS-CoV-2, numerous mutations with known negative impact on diagnostic assays have been identified. In early 2021, we identified four samples positive for SARS-CoV-2 with a nucleocapsid (N) gene drop out on Cepheid Xpert® Xpress SARS-CoV-2 assay. Sequencing revealed a single common mutation in the N gene C29200T. Spatiotemporal analysis showed that the mutation was found in at least six different Canadian provinces from May 2020 until May 2021. Phylogenetic analysis showed that this mutation arose multiple times in Canadian samples and is present in six different variants of interest and of concern. The Cepheid testing platform is commonly used in Canada including in remote regions. As such, the existence of N gene mutation dropouts required further investigation. While commercial SARS-CoV-2 molecular detection assays have contributed immensely to the response effort, many vendors are reluctant to make primer/probe sequences publicly available. Proprietary primer/probe sequences create diagnostic 'blind spots' for global SARS-CoV-2 sequence monitoring and limits the ability to detect and track the presence and prevalence of diagnostic escape mutations. We hope that our industry partners will seriously consider making primer/probe sequences available, so that diagnostic escape mutants can be identified promptly and responded to appropriately to maintain diagnostic accuracy.