Multiplex Quantitative Polymerase Chain Reaction Diagnostic Test for SARS-CoV-2 and Influenza A/B Viruses

Steve F C Hawkins; Paul C Guest

doi:10.1007/978-1-0716-2395-4_4

Multiplex Quantitative Polymerase Chain Reaction Diagnostic Test for SARS-CoV-2 and Influenza A/B Viruses

Methods Mol Biol. 2022:2511:53-65. doi: 10.1007/978-1-0716-2395-4_4.

Authors

Steve F C Hawkins¹, Paul C Guest²

Affiliations

¹ Meridian Bioscience, Unit 16, The Edge Business Centre, London, UK. shawkins@bioline.com.
² Laboratory of Neuroproteomics, Department of Biochemistry and Tissue Biology, Institute of Biology, University of Campinas (UNICAMP), Campinas, Brazil.

PMID: 35838951
DOI: 10.1007/978-1-0716-2395-4_4

Abstract

COVID-19 disease caused by the novel SARS-CoV-2 virus represents a new challenge for healthcare systems. The molecular confirmation of infection is crucial to guide public health decision-making. This task could be made more difficult during the next influenza season. Thus, a rapid and user-friendly diagnostic test to discriminate SARS-CoV-2 from influenza viruses is urgently needed. Here, we present a multiplex quantitative polymerase chain reaction (qPCR) assay capable of distinguishing SARS-CoV-2 from influenza A and B cases. This assay benefits from the use of an inhibitor tolerant PCR mix which obviates the need for the rate-limiting extraction step, allowing for a more rapid and accurate analysis.

Keywords: COVID-19; Extraction; Influenza; Inhibitor-tolerant; Quantitation; SARS-CoV-2; qPCR.

MeSH terms

COVID-19* / diagnosis
Diagnostic Tests, Routine
Herpesvirus 1, Cercopithecine*
Humans
Influenza A virus* / genetics
Influenza B virus
Influenza, Human* / diagnosis
Multiplex Polymerase Chain Reaction
Real-Time Polymerase Chain Reaction
SARS-CoV-2 / genetics
Sensitivity and Specificity