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Abstract

The QuickGene-810 Nucleic Acid Isolation System is a semi-automated extraction platform which may be used for RNA extraction. New methods were required to support the rapid increase in respiratory virus testing during the SARS-CoV-2 pandemic. The aim of this study was to assess SARS-CoV-2 RNA extraction using the QuickGene-810 kit compared to the EZ1 Advanced Extraction Platform for use on the AusDiagnostics SARS-CoV-2, Influenza and RSV 8-well RT-PCR assay. Qualitative results from all clinical samples were concordant between the QuickGene-810 and the EZ1 extraction methods, demonstrating that the QuickGene-810 kit is suitable for use in pathogen diagnostics. However, there was an average difference of approximately two cycles between the cycle threshold (Ct) values for both SARS-CoV-2 targets, suggesting that the EZ1 kit yields a higher concentration of nucleic acid extract, possibly related to its use of carrier RNA and/or smaller elution volume, which infers the possibility of false negative results for samples with very low viral loads.

  • This is an open-access article distributed under the terms of the Creative Commons Attribution License.
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/content/journal/acmi/10.1099/acmi.0.000353
2022-05-27
2024-12-08
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References

  1. Phan TS, Stephenson R, Cai T, Andacic N, McKew G. A comparison of SARS-cov-2 RNA extraction with the quickgene-810 nucleic acid isolation system compared to the EZ1 advanced DSP virus kit Figshare 2020 DOI: 10.6084/m9.figshare.19316048
    [Google Scholar]
  2. Mercer TR, Salit M. Testing at scale during the COVID-19 pandemic. Nat Rev Genet 2021; 22:415–426 [View Article] [PubMed]
    [Google Scholar]
  3. American Society for Microbiology Supply Shortages Impacting COVID-19 and Non-COVID tTesting. Washington, DC: ASM; 2021 https://asm.org/Articles/2020/September/Clinical-Microbiology-Supply-Shortage-Collecti-1
  4. He H, Li R, Chen Y, Pan P, Tong W et al. Integrated DNA and RNA extraction using magnetic beads from viral pathogens causing acute respiratory infections. Sci Rep 2017; 7:45199 [View Article] [PubMed]
    [Google Scholar]
  5. Phan T. The method verification of the 8 well Ausdiagnostics SARS CoV2 assay. In Concord Hospital Microbiology Concord, Australia: NSW Health Pathology; 2020
    [Google Scholar]
  6. AusDiagnostics AusDiagnostics High-Plex 24 System REF 91501. Instructions for Use Mascot, Australia: AusDiagnostics; 2019 pp 1–44
    [Google Scholar]
  7. Hiroi S, Kubota-Koketsu R, Sasaki T, Morikawa S, Motomura K et al. Infectivity assay for detection of SARS-CoV-2 in samples from patients with COVID-19. J Med Virol 2021; 93:5917–5923 [View Article] [PubMed]
    [Google Scholar]
  8. Yamada S, Fukushi S, Kinoshita H, Ohnishi M, Suzuki T et al. Assessment of SARS-CoV-2 infectivity of upper respiratory specimens from COVID-19 patients by virus isolation using VeroE6/TMPRSS2 cells. BMJ Open Respir Res 2021; 8:e000830 [View Article] [PubMed]
    [Google Scholar]
  9. Paton TF, Marr I, O’Keefe Z, Inglis TJJ. Development, deployment and in-field demonstration of mobile coronavirus SARS-CoV-2 Nucleic acid amplification test. J Med Microbiol 2021; 70:001346 [View Article] [PubMed]
    [Google Scholar]
  10. Eichhoff OM, Bellini E, Lienhard R, Stark WJ, Bechtold P et al. Comparison of RNA extraction methods for the detection ofSARS-CoV-2 by RT-PCR. 2020.08.13.20172494 medRxiv [Preprint] 2020 [View Article]
    [Google Scholar]
  11. Lim H-J, Jung H-S, Park M-Y, Baek Y-H, Kannappan B et al. Evaluation of three automated extraction systems for the detection of SARS-CoV-2 from clinical respiratory specimens. Life (Basel) 2022; 12:68 [View Article] [PubMed]
    [Google Scholar]
  12. Ransom EM, Potter RF, Wallace MA, Mitchell KF, Yarbrough ML et al. Comparison of extraction methods and thermocyclers for SARS-CoV-2 molecular detection using clinical specimens. J Clin Microbiol 2020; 58:e01622-20 [View Article] [PubMed]
    [Google Scholar]
  13. Wu S, Shi X, Chen Q, Jiang Y, Zuo L et al. Comparative evaluation of six nucleic acid amplification kits for SARS-CoV-2 RNA detection. Ann Clin Microbiol Antimicrob 2021; 20:38 [View Article] [PubMed]
    [Google Scholar]
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