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Updated: Nov 16, 2025

Large-Scale SARS-CoV-2 Testing Utilizing Saliva and Transposition Sample Pooling
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Streamlining SARS-CoV-2 confirmatory testing to reduce false positive results.

Michael J Wilson1, Dominic Sparkes2, Chloe Myers3

  • 1Department of Virology, Cambridge University Hospitals NHS Foundation Trust, Cambridge, UK; Clinical Microbiology & Public Health Laboratory, Public Health England, Cambridge, UK.

Journal of Clinical Virology : the Official Publication of the Pan American Society for Clinical Virology
|February 19, 2021
PubMed
Summary

Confirmatory testing for SARS-CoV-2 (severe acute respiratory syndrome coronavirus 2) can reduce false positives but increases workload. A Relative Light Unit (RLU) threshold of 1000 efficiently identifies samples needing confirmation, improving accuracy and lab efficiency.

Keywords:
COVID-19Confirmatory testingFalse positiveLaboratory diagnosisSARS-CoV-2

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Area of Science:

  • Clinical diagnostics
  • Virology
  • Laboratory medicine

Background:

  • Confirmatory testing for SARS-CoV-2 is crucial for accuracy.
  • Current methods increase laboratory workload and turnaround time.
  • Optimizing confirmatory testing is necessary during periods of high case numbers.

Purpose of the Study:

  • To analyze the laboratory's confirmatory testing pathway for SARS-CoV-2.
  • To establish a refined approach for confirmatory testing.
  • To balance accuracy with laboratory efficiency and reduce workload.

Main Methods:

  • Analyzed confirmatory testing data over a 10-week low prevalence period.
  • Measured turnaround time for confirmatory tests.
  • Identified a Relative Light Unit (RLU) threshold for cut-off determination.

Main Results:

  • 30% of newly positive SARS-CoV-2 samples were false positives.
  • Confirmatory testing increased turnaround time by 17 hours.
  • A RLU threshold of 1000 significantly differentiated true positives (98% confirmation) from potential false positives (32% confirmation).

Conclusions:

  • Establishing an RLU threshold streamlines confirmatory testing for SARS-CoV-2.
  • This approach focuses confirmatory testing on high-risk samples, improving efficiency.
  • Laboratories should develop similar RLU thresholds for their assays to enhance accuracy and efficiency during high case loads.