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Exploring beyond the limit: How comparative stochastic performance affects retesting outcomes in six commercial SARS
Hiu Tat Chan1,2, Marco H T Keung1, Ivy Nguyen1
1VCS Pathology, Australian Centre for the Prevention of Cervical Cancer, 265 Faraday Street, Carlton South, VIC 3053, Australia.
Objectives:
To examine the comparative stochasticity profile of six commercial SARS-CoV-2 nucleic acid amplification tests (NAATs) and how this may affect retesting paradigms.
Methods:
Commercial quality control (QC) material was serially diluted in viral transport media to create a panel covering 10-10,000 copies/ml. The panel was tested across six commercial NAATs. A subset of high cycle threshold results was retested on a rapid PCR assay to simulate retesting protocols commonly used to discriminate false positives.
Results:
Performance beyond the LOD differed among assays, with three types of stochasticity profiles observed. The ability of the rapid PCR assay to reproduce a true weak positive specimen was restricted to its own stochastic performance at the corresponding viral concentration.
Conclusion:
Stochastic performance of various NAATs overlap across low viral concentrations and affect retesting outcomes. Relying on retesting alone to discriminate false positives risk missing true positives even when a more sensitive assay is deployed for confirmatory testing.
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