tetano
Editor, Senior Moderator
Sci Rep
. 2022 Mar 3;12(1):3480.
doi: 10.1038/s41598-022-07152-0.
Clinical validation of a multiplex PCR-based detection assay using saliva or nasopharyngeal samples for SARS-Cov-2, influenza A and B
Nikhil S Sahajpal[SUP] 1 [/SUP], Ashis K Mondal[SUP] 1 [/SUP], Sudha Ananth[SUP] 1 [/SUP], Allan Njau[SUP] 2 [/SUP], Kimya Jones[SUP] 1 [/SUP], Pankaj Ahluwalia[SUP] 1 [/SUP], Eesha Oza[SUP] 1 [/SUP], Ted M Ross[SUP] 3 [/SUP], Vamsi Kota[SUP] 4 [/SUP], Arvind Kothandaraman[SUP] 5 [/SUP], Sadanand Fulzele[SUP] 6 [/SUP], Madhuri Hegde[SUP] 5 [/SUP], Alka Chaubey[SUP] 1 7 [/SUP], Amyn M Rojiani[SUP] 8 [/SUP], Ravindra Kolhe[SUP] 9 [/SUP]
Affiliations
Abstract
The COVID-19 pandemic has resulted in significant diversion of human and material resources to COVID-19 diagnostics, to the extent that influenza viruses and co-infection in COVID-19 patients remains undocumented and pose serious public-health consequences. We optimized and validated a highly sensitive RT-PCR based multiplex-assay for the detection of SARS-CoV-2, influenza A and B viruses in a single-test. This study evaluated clinical specimens (n = 1411), 1019 saliva and 392 nasopharyngeal swab (NPS), tested using two-assays: FDA-EUA approved SARS-CoV-2 assay that targets N and ORF1ab gene, and the PKamp-RT-PCR based assay that targets SARS-CoV-2, influenza viruses A and B. Of the 1019 saliva samples, 17.0% (174/1019) tested positive for SARS-CoV-2 using either assay. The detection rate for SARS-CoV-2 was higher with the multiplex assay compared to SARS-specific assay [91.9% (160/174) vs. 87.9% (153/174)], respectively. Of the 392 NPS samples, 10.4% (41/392) tested positive for SARS-CoV-2 using either assay. The detection rate for SARS-CoV-2 was higher with the multiplex assay compared to SARS-specific assay [97.5% (40/41) vs. 92.1% (39/41)], respectively. This study presents clinical validation of a multiplex-PCR assay for testing SARS-CoV-2, influenza A and B viruses, using NPS and saliva samples, and demonstrates the feasibility of implementing the assay without disrupting the existing laboratory workflow.
. 2022 Mar 3;12(1):3480.
doi: 10.1038/s41598-022-07152-0.
Clinical validation of a multiplex PCR-based detection assay using saliva or nasopharyngeal samples for SARS-Cov-2, influenza A and B
Nikhil S Sahajpal[SUP] 1 [/SUP], Ashis K Mondal[SUP] 1 [/SUP], Sudha Ananth[SUP] 1 [/SUP], Allan Njau[SUP] 2 [/SUP], Kimya Jones[SUP] 1 [/SUP], Pankaj Ahluwalia[SUP] 1 [/SUP], Eesha Oza[SUP] 1 [/SUP], Ted M Ross[SUP] 3 [/SUP], Vamsi Kota[SUP] 4 [/SUP], Arvind Kothandaraman[SUP] 5 [/SUP], Sadanand Fulzele[SUP] 6 [/SUP], Madhuri Hegde[SUP] 5 [/SUP], Alka Chaubey[SUP] 1 7 [/SUP], Amyn M Rojiani[SUP] 8 [/SUP], Ravindra Kolhe[SUP] 9 [/SUP]
Affiliations
- PMID: 35241679
- DOI: 10.1038/s41598-022-07152-0
Abstract
The COVID-19 pandemic has resulted in significant diversion of human and material resources to COVID-19 diagnostics, to the extent that influenza viruses and co-infection in COVID-19 patients remains undocumented and pose serious public-health consequences. We optimized and validated a highly sensitive RT-PCR based multiplex-assay for the detection of SARS-CoV-2, influenza A and B viruses in a single-test. This study evaluated clinical specimens (n = 1411), 1019 saliva and 392 nasopharyngeal swab (NPS), tested using two-assays: FDA-EUA approved SARS-CoV-2 assay that targets N and ORF1ab gene, and the PKamp-RT-PCR based assay that targets SARS-CoV-2, influenza viruses A and B. Of the 1019 saliva samples, 17.0% (174/1019) tested positive for SARS-CoV-2 using either assay. The detection rate for SARS-CoV-2 was higher with the multiplex assay compared to SARS-specific assay [91.9% (160/174) vs. 87.9% (153/174)], respectively. Of the 392 NPS samples, 10.4% (41/392) tested positive for SARS-CoV-2 using either assay. The detection rate for SARS-CoV-2 was higher with the multiplex assay compared to SARS-specific assay [97.5% (40/41) vs. 92.1% (39/41)], respectively. This study presents clinical validation of a multiplex-PCR assay for testing SARS-CoV-2, influenza A and B viruses, using NPS and saliva samples, and demonstrates the feasibility of implementing the assay without disrupting the existing laboratory workflow.