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Volume 4, Issue 3

Comparative evaluation of Panther Fusion and real-time PCR for detection of in spiked human blood Open Access

Abstract

Melioidosis is an infection that most commonly presents with bacteraemia. Culture-based laboratory methods can result in a significant delay to organism identification. Molecular diagnostic techniques have a high sensitivity and rapid time to diagnosis. A decreased time to diagnosis is likely to improve patient outcomes.

To compare the Panther Fusion automated molecular instrument to an in-house method for the detection of directly from spiked human whole-blood samples.

The in-house method detected 11/12 (92 %) samples with a concentration of 2.5–4.5×10 c.f.u. ml. The Panther was less reliable, detecting only 8/14 (75 %) samples with a similar bacterial concentration. The Panther was able to detect 12/12 (100 %) spiked blood culture-positive samples.

The direct detection of from patient blood on presentation to a healthcare facility will significantly decrease time to diagnosis. We describe an in-house real-time PCR method with the lowest reported limit of detection to date. Due to lower sensitivity, the Panther Fusion would be best used as a diagnostic method directly from a positive blood culture.

  • Received:
  • Accepted:
  • Published Online:
Keyword(s): bacteraemia , blood culture , Burkholderia pseudomallei , melioidosis , molecular detection , PCR and TTS1
Funding
This study was supported by the:
  • Pathology Queensland (Award SERC 6096)
    • Principle Award Recipient: IanGassiep
  • Hologic
    • Principle Award Recipient: IanGassiep
© 2022 The Authors
  • This is an open-access article distributed under the terms of the Creative Commons Attribution License. This article was made open access via a Publish and Read agreement between the Microbiology Society and the corresponding author’s institution.
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2022-03-21
2024-04-26
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References

  1. Gassiep I, Armstrong M, Norton R. Human Melioidosis. Clin Microbiol Rev 2020; 33:e00006–00019 [View Article] [PubMed]
     [Google Scholar]
  2. Limmathurotsakul D, Golding N, Dance DA, Messina JP, Pigott DM et al. Predicted global distribution of Burkholderia pseudomallei and burden of melioidosis. Nat Microbiol 2016; 1:15008 [View Article] [PubMed]
     [Google Scholar]
  3. Birnie E, Virk HS, Savelkoel J, Spijker R, Bertherat E et al. Global burden of melioidosis in 2015: a systematic review and data synthesis. Lancet Infect Dis 2019; 19:892–902 [View Article] [PubMed]
     [Google Scholar]
  4. Hantrakun V, Kongyu S, Klaytong P, Rongsumlee S, Day NPJ et al. Clinical Epidemiology of 7126 Melioidosis Patients in Thailand and the Implications for a National Notifiable Diseases Surveillance System. Open Forum Infect Dis 2019; 6:fz498 [View Article] [PubMed]
     [Google Scholar]
  5. Currie BJ, Mayo M, Ward LM, Kaestli M, Meumann EM et al. The Darwin Prospective Melioidosis Study: a 30-year prospective, observational investigation. Lancet Infect Dis 2021; 21:1737–1746 [View Article] [PubMed]
     [Google Scholar]
  6. Gassiep I, Ganeshalingam V, Chatfield MD, Harris PNA, Norton RE. The epidemiology of melioidosis in Townsville, Australia. Trans R Soc Trop Med Hyg 2021trab125 [View Article] [PubMed]
     [Google Scholar]
  7. Limmathurotsakul D, Jamsen K, Arayawichanont A, Simpson JA, White LJ et al. Defining the true sensitivity of culture for the diagnosis of melioidosis using Bayesian latent class models. PLoS One 2010; 5:e12485 [View Article] [PubMed]
     [Google Scholar]
  8. Lew AE, Desmarchelier PM. Detection of Pseudomonas pseudomallei by PCR and hybridization. J Clin Microbiol 1994; 32:1326–1332 [View Article] [PubMed]
     [Google Scholar]
  9. Gassiep I, Burnard D, Bauer MJ, Norton RE, Harris PN. Diagnosis of melioidosis: the role of molecular techniques. Future Microbiol 2021; 16:271–288 [View Article] [PubMed]
     [Google Scholar]
  10. Tiangpitayakorn C, Songsivilai S, Piyasangthong N, Dharakul T. Speed of detection of Burkholderia pseudomallei in blood cultures and its correlation with the clinical outcome. Am J Trop Med Hyg 1997; 57:96–99 [View Article] [PubMed]
     [Google Scholar]
  11. Ombelet S, Peeters M, Phe C, Tsoumanis A, Kham C et al. Nonautomated blood cultures in a low-resource setting: optimizing the timing of blind subculture. Am J Trop Med Hyg 2020; 104:612–621 [View Article] [PubMed]
     [Google Scholar]
  12. Winstanley C, Hales BA, Hart CA. Evidence for the presence in Burkholderia pseudomallei of a type III secretion system-associated gene cluster. J Med Microbiol 1999; 48:649–656 [View Article] [PubMed]
     [Google Scholar]
  13. Kaestli M, Richardson LJ, Colman RE, Tuanyok A, Price EP et al. Comparison of TaqMan PCR assays for detection of the melioidosis agent Burkholderia pseudomallei in clinical specimens. J Clin Microbiol 2012; 50:2059–2062 [View Article] [PubMed]
     [Google Scholar]
  14. Novak RT, Glass MB, Gee JE, Gal D, Mayo MJ et al. Development and evaluation of a real-time PCR assay targeting the type III secretion system of Burkholderia pseudomallei. J Clin Microbiol 2006; 44:85–90 [View Article] [PubMed]
     [Google Scholar]
  15. Meumann EM, Novak RT, Gal D, Kaestli ME, Mayo M et al. Clinical evaluation of a type III secretion system real-time PCR assay for diagnosing melioidosis. J Clin Microbiol 2006; 44:3028–3030 [View Article] [PubMed]
     [Google Scholar]
  16. Chantratita N, Meumann E, Thanwisai A, Limmathurotsakul D, Wuthiekanun V et al. Loop-mediated isothermal amplification method targeting the TTS1 gene cluster for detection of Burkholderia pseudomallei and diagnosis of melioidosis. J Clin Microbiol 2008; 46:568–573 [View Article] [PubMed]
     [Google Scholar]
  17. Podnecky NL, Elrod MG, Newton BR, Dauphin LA, Shi J et al. Comparison of DNA extraction kits for detection of Burkholderia pseudomallei in spiked human whole blood using real-time PCR. PLoS One 2013; 8:e58032 [View Article] [PubMed]
     [Google Scholar]
  18. Stellrecht KA, Cimino JL, Maceira VP. The panther fusion system with open access functionality for laboratory-developed tests for influenza A virus subtyping. J Clin Microbiol 2020; 58:e00188-20 [View Article] [PubMed]
     [Google Scholar]
  19. Lieberman JA, Pepper G, Naccache SN, Huang M-L, Jerome KR et al. Comparison of Commercially Available and Laboratory-Developed Assays for In Vitro Detection of SARS-CoV-2 in Clinical Laboratories. J Clin Microbiol 2020; 58:e00821–00820 [View Article] [PubMed]
     [Google Scholar]
  20. Frontzek A, Aretzweiler G, Winkens D, Duncan D, Marlowe EM. High-volume workflow and performance comparisons for Chlamydia trachomatis and Neisseria gonorrhoeae testing using automated molecular platforms. BMC Infect Dis 2019; 19:797 [View Article] [PubMed]
     [Google Scholar]
  21. Messacar K, Parker SK, Todd JK, Dominguez SR. Implementation of rapid molecular infectious disease diagnostics: the role of diagnostic and antimicrobial stewardship. J Clin Microbiol 2017; 55:715–723 [View Article] [PubMed]
     [Google Scholar]
  22. Gassiep I, Armstrong M, Norton RE. Identification of Burkholderia pseudomallei by Use of the Vitek Mass Spectrometer. J Clin Microbiol 2019; 57:e00081-19 [View Article] [PubMed]
     [Google Scholar]
  23. Watthanaworawit W, Roberts T, Hopkins J, Gassiep I, Norton R et al. A multi-country study using MALDI-TOF mass spectrometry for rapid identification of Burkholderia pseudomallei. BMC Microbiol 2021; 21:213 [View Article] [PubMed]
     [Google Scholar]
  24. Wuthiekanun V, Limmathurotsakul D, Wongsuvan G, Chierakul W, Teerawattanasook N et al. Quantitation of B. Pseudomallei in clinical samples. Am J Trop Med Hyg 2007; 77:812–813 [View Article] [PubMed]
     [Google Scholar]
  25. Kim SC, Lee S, Kim S, Cho OH, Park H et al. Comparison of Clinical Performance Between BacT/Alert Virtuo and BacT/Alert 3D Blood Culture Systems. Ann Lab Med 2019; 39:278–283 [View Article] [PubMed]
     [Google Scholar]
  26. Burd EM. Validation of laboratory-developed molecular assays for infectious diseases. Clin Microbiol Rev 2010; 23:550–576 [View Article] [PubMed]
     [Google Scholar]
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Comparative evaluation of Panther Fusion and real-time PCR for detection of Burkholderia pseudomallei in spiked human blood
Access Microbiology 4, 000333 (2022); https://doi.org/10.1099/acmi.0.000333
/content/journal/acmi/10.1099/acmi.0.000333
/content/journal/acmi/10.1099/acmi.0.000333
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