Validation of a molecular assay to detect SARS-CoV-2 in saliva

Janet L. Pitman, Arthur J. Morris, Stephen Grice, Joseph T. Walsh, Leyi Wang, Martin D. Burke, Amanda Dixon-McIver

Research output: Contribution to journalArticlepeer-review

Abstract

AIM: To validate a reverse transcriptase-quantitative polymerase chain reaction (RT-qPCR) assay to detect SARS-CoV-2 in saliva in two independent Aotearoa New Zealand laboratories. METHODS: An RT-qPCR assay developed at University of Illinois Urbana-Champaign, USA, was validated in two New Zealand laboratories. Analytical measures, such as limit of detection (LOD) and cross-reactivity, were performed. One hundred and forty-seven saliva samples, each paired with a contemporaneously collected nasal swab, mainly of nasopharyngeal origin, were received. Positive (N=33) and negative (N=114) samples were tested blindly in each laboratory. Diagnostic sensitivity and specificity were then calculated. RESULTS: The LOD was <0.75 copy per µL and no cross-reactivity with MERS-CoV was detected. There was complete concordance between laboratories for all saliva samples with the quantification cycle values for all three genes in close agreement. Saliva had 98.7% concordance with paired nasal samples: and a sensitivity, specificity and accuracy of 97.0%, 99.1% and 99.1%, respectively. CONCLUSION: This saliva RT-qPCR assay produces reproducible results with a low LOD. High sensitivity and specificity make it a reliable option for SARS-CoV-2 testing, including for asymptomatic people requiring regular screening.

Original languageEnglish (US)
Pages (from-to)14-27
Number of pages14
JournalNew Zealand Medical Journal
Volume134
Issue number1547
StatePublished - Dec 17 2021

ASJC Scopus subject areas

  • Medicine(all)

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