Effect of not vortexing nasopharyngeal and throat swabs on SARS-CoV-2 nucleic acid detection: A pilot study

• Published in: Journal of virological methods 2022-03, Vol.301, p.114468-114468, Article 114468
• Main Authors: Kanji, Jamil N., Pabbaraju, Kanti, Wong, Anita, Beitku, Candace, Deo, Ashwin, Kailey, Seema, Ma, Raymond, Tipples, Graham, Zelyas, Nathan
• Format: Article
• Language: English
• Subjects: automation; COVID-19; COVID-19 Testing; gene amplification; Humans; Nasopharynx; PCR; Pharynx; Pilot Projects; Protocols; Qualitative; RNA; RNA, Viral - genetics; Sample processing; SARS-CoV-2; Severe acute respiratory syndrome coronavirus 2; Specimen Handling - methods; throat; viruses; Vortex
• ISSN: 0166-0934; 1879-0984
• DOI: 10.1016/j.jviromet.2022.114468

•High-throughput COVID-19 testing typically involves the vortexing of swab samples.•In this pilot study, omitting this step did not impact SARS-CoV-2 detection.•This was observed consistently across eight separate SARS-CoV-2 nucleic acid tests.•Such pre-analytical improvements streamline testing and reduce contamination risk. The processing of swabs for respiratory virus detection involves vortexing while still in the viral transport medium (VTM). The effect of not vortexing swabs prior to analysis has not been studied extensively for SARS-CoV-2 detection, and presents an opportunity to improve pre-analytic laboratory workflow. We aimed to assess the impact of not vortexing nasopharyngeal/throat swabs submitted in VTM for SARS-CoV-2 testing. To assess the impact of not vortexing swabs, 277 swab samples were tested for SARS-CoV-2 RNA in paired vortexed and non-vortexed aliquots using eight routine nucleic acid amplification assays. We compared the qualitative (positive/negative) and semi-quantitative (cycle threshold, Ct) results. Following discordant analysis, all but one non-vortexed sample had the same qualitative result as the vortexed sample. 27.4 % of samples were SARS-CoV-2 positive. Comparison of Ct values revealed an apparent reduction in human cellular nucleic acid in the non-vortexed samples (mean Ct values of 24.0 and 26.5 for vortexed and non-vortexed samples, respectively, p < 0.0001) and increased Ct values for non-vortexed samples using a laboratory-developed SARS-CoV-2 assay (mean Ct values of 4.1 and 4.2 for vortexed and non-vortexed samples, respectively; p < 0.0001), but this was not observed for a more automated commercial SARS-CoV-2 assay (mean Ct values of 15.2 for both vortexed and non-vortexed samples, respectively; p = 0.68). While vortexing swabs appears to improve the recovery of cellular material, it does not have an appreciable impact on the qualitative sensitivity of SARS-CoV-2 nucleic acid tests, which may support omission of this step and simplification of front-end sample processing.