Assessing the comparability of cycle threshold values derived from five external quality assessment rounds for omicron nucleic acid testing

Abstract

Abstract Background A variety of open-system real-time reverse transcriptase polymerase chain reaction (RT-PCR) assays for several acute respiratory syndrome coronavirus 2 are currently in use. This study aimed to ensure the quality of omicron nucleic acid testing and to assess the comparability of cycle threshold (Ct) values derived from RT-PCR. Methods Five external quality assessment (EQA) rounds using the omicron virus-like particles were organized between February 2022 and June 2022. Results A total of 1401 qualitative EQA reports have been collected. The overall positive percentage agreement was 99.72%, the negative percentage agreement was 99.75%, and the percent agreement was 99.73%. This study observed a significant variance in Ct values derived from different test systems. There was a wide heterogeneity in PCR efficiency among different RT-PCR kits and inter-laboratories. Conclusion There was strong concordance among laboratories performing qualitative omicron nucleic acid testing. Ct values from qualitative RT-PCR tests should not be used for clinical or epidemiological decision-making to avoid the potential for misinterpretation of the results.

Keywords

• SARS-CoV-2
• Omicron
• RT-PCR
• Cycle threshold
• Qualitative test
• Comparability