1887

Abstract

Laboratories worldwide are facing high demand for molecular testing during the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic, which might be further aggravated by the upcoming influenza season in the northern hemisphere.

Given that the symptoms of influenza are largely indistinguishable from those of coronavirus disease 2019 (COVID-19), both SARS-CoV-2 and the influenza viruses require concurrent testing by RT-PCR in patients presenting with symptoms of respiratory tract infection.

We adapted and evaluated a laboratory-developed multiplex RT-PCR assay for simultaneous detection of SARS-CoV-2 (dual target), influenza A and influenza B (SC2/InflA/InflB-UCT) on a fully automated high-throughput system (cobas6800).

Analytical performance was assessed by serial dilution of quantified reference material and cell culture stocks in transport medium, including pretreatment for chemical inactivation. For clinical evaluation, residual portions of 164 predetermined patient samples containing SARS-CoV-2 (=52), influenza A (=43) or influenza B (=19), as well as a set of negative samples, were subjected to the novel multiplex assay.

The assay demonstrated comparable analytical performance to currently available commercial tests, with limits of detection of 94.9 cp ml for SARS-CoV-2, 14.6 cp ml for influenza A and 422.3 cp ml for influenza B. Clinical evaluation showed excellent agreement with the comparator assays (sensitivity of 98.1, 97.7 and 100 % for Sars-CoV-2 and influenza A and B, respectively).

The SC2/InflA/InflB-UCT allows for efficient high-throughput testing for all three pathogens and thus provides streamlined diagnostics while conserving resources during the influenza season.

  • This is an open-access article distributed under the terms of the Creative Commons Attribution NonCommercial License. The Microbiology Society waived the open access fees for this article.
Loading

Article metrics loading...

/content/journal/jmm/10.1099/jmm.0.001295
2021-01-06
2024-10-10
Loading full text...

Full text loading...

/deliver/fulltext/jmm/70/2/jmm001295.html?itemId=/content/journal/jmm/10.1099/jmm.0.001295&mimeType=html&fmt=ahah

References

  1. Wu Z, McGoogan JM. Characteristics of and important lessons from the coronavirus disease 2019 (COVID-19) outbreak in china: summary of a report of 72 314 cases from the chinese center for disease control and prevention. JAMA 2020; 323:1239-1242 [View Article][PubMed]
    [Google Scholar]
  2. Stokes EK, Zambrano LD, Anderson KN, Marder EP, Raz KM et al. Coronavirus disease 2019 case surveillance - United States, January 22-May 30, 2020. MMWR Morb Mortal Wkly Rep 2020; 69:759–765 [View Article][PubMed]
    [Google Scholar]
  3. Eigner U, Reucher S, Hefner N, Staffa-Peichl S, Kolb M et al. Clinical evaluation of multiplex RT-PCR assays for the detection of influenza A/B and respiratory syncytial virus using a high throughput system. J Virol Methods 2019; 269:49–54 [View Article][PubMed]
    [Google Scholar]
  4. Pfefferle S, Reucher S, Nörz D, Lütgehetmann M. Evaluation of a quantitative RT-PCR assay for the detection of the emerging coronavirus SARS-CoV-2 using a high throughput system. Euro Surveill 2020; 25:2000152 [View Article][PubMed]
    [Google Scholar]
  5. Poljak M, Korva M, Knap Gašper N, Fujs Komloš K, Sagadin M et al. Clinical evaluation of the COBAS SARS-CoV-2 test and a diagnostic platform switch during 48 hours in the midst of the COVID-19 pandemic. J Clin Microbiol 2020; 58: [View Article]
    [Google Scholar]
  6. Corman VM, Landt O, Kaiser M, Molenkamp R, Meijer A et al. Detection of 2019 novel coronavirus (2019-nCoV) by real-time RT-PCR. Euro Surveill 2020; 25: [View Article]
    [Google Scholar]
  7. Chan JF-W, Yip CC-Y, To KK-W, Tang TH-C, Wong SC-Y et al. Improved molecular diagnosis of COVID-19 by the novel, highly sensitive and specific COVID-19-RdRp/Hel Real-Time reverse transcription-PCR assay validated In Vitro and with clinical specimens. J Clin Microbiol 2020; 58:e00310–00320 [View Article][PubMed]
    [Google Scholar]
  8. WHO Who information for the molecular detection of influenza viruses. WHO Website 2017
    [Google Scholar]
  9. Terrier O, Josset L, Textoris J, Marcel V, Cartet G et al. Cellular transcriptional profiling in human lung epithelial cells infected by different subtypes of influenza A viruses reveals an overall down-regulation of the host p53 pathway. Virol J 2011; 8:285-285 [View Article][PubMed]
    [Google Scholar]
  10. CDC Research use only CDC influenza SARS-CoV-2 (flu SC2) multiplex assay real-time RT-PCR primers and probes. CDC Website 2020
    [Google Scholar]
  11. Pfefferle S, Huang J, Nörz D, Indenbirken D, Lütgehetmann M et al. Complete genome sequence of a SARS-CoV-2 strain isolated in northern Germany. Microbiol Resour Announc 2020; 9:e00520–20 [View Article][PubMed]
    [Google Scholar]
  12. Nörz D, Frontzek A, Eigner U, Oestereich L, Wichmann D, Fischer N et al. Pushing beyond specifications: evaluation of linearity and clinical performance of the COBAS 6800/8800 SARS-CoV-2 RT-PCR assay for reliable quantification in blood and other materials outside recommendations. J Clin Virol 2020; 132:104650 [View Article][PubMed]
    [Google Scholar]
  13. Vogels CBF, Brito AF, Wyllie AL, Fauver JR, Ott IM et al. Analytical sensitivity and efficiency comparisons of SARS-CoV-2 RT-qPCR primer-probe sets. Nat Microbiol 2020; 5:1299–1305 [View Article][PubMed]
    [Google Scholar]
/content/journal/jmm/10.1099/jmm.0.001295
Loading
/content/journal/jmm/10.1099/jmm.0.001295
Loading

Data & Media loading...

Supplements

Supplementary material 1

PDF
This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error