1887

Abstract

Molecular diagnostic assays can accelerate the diagnosis of fungal infections and subsequently improve patient outcomes. In particular, the detection of infections due to Mucorales is still challenging for laboratories and physicians. The aim of this study was to evaluate a probe-based Mucorales-specific real-time PCR assay (Muc18S) using tissue and serum samples from patients suffering from invasive mucormycosis (IMM). This assay can detect a broad range of clinically relevant Mucorales species and can be used to complement existing diagnostic tests or to screen high-risk patients. An advantage of the Muc18S assay is that it exclusively detects Mucorales species allowing the diagnosis of Mucorales DNA without sequencing within a few hours. In paraffin-embedded tissue samples this PCR-based method allowed rapid identification of Mucorales in comparison with standard methods and showed 91 % sensitivity in the IMM tissue samples. We also evaluated serum samples, an easily accessible material, from patients at risk from IMM. Mucorales DNA was detected in all patients with probable/proven IMM (100 %) and in 29 % of the possible cases. Detection of IMM in serum could enable an earlier diagnosis (up to 21 days) than current methods including tissue samples, which were gained mainly post-mortem. A screening strategy for high-risk patients, which would enable targeted treatment to improve patient outcomes, is therefore possible.

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/content/journal/jmm/10.1099/jmm.0.000375
2016-12-16
2021-10-16
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References

  1. Alanio A., Garcia-Hermoso D., Mercier-Delarue S., Lanternier F., Gits-Muselli M., Menotti J., Denis B., Bergeron A., Legrand M. et al. 2015; Molecular identification of Mucorales in human tissues: contribution of PCR electrospray-ionization mass spectrometry. Clin Microbiol Infect 21:594.e1594.e5 [View Article]
    [Google Scholar]
  2. Antinori S., Nebuloni M., Magni C., Fasan M., Adorni F., Viola A., Corbellino M., Galli M., Vago G. et al. 2009; Trends in the postmortem diagnosis of opportunistic invasive fungal infections in patients with AIDS: a retrospective study of 1,630 autopsies performed between 1984 and 2002. Am J Clin Pathol 132:221227 [View Article][PubMed]
    [Google Scholar]
  3. Armitage P., Berry G., Matthews J. N. S. 2002 Statistical Methods in Medical Research Oxford: Blackwell Scientific;
    [Google Scholar]
  4. Auberger J., Lass-Flörl C., Aigner M., Clausen J., Gastl G., Nachbaur D. 2012; Invasive fungal breakthrough infections, fungal colonization and emergence of resistant strains in high-risk patients receiving antifungal prophylaxis with posaconazole: real-life data from a single-centre institutional retrospective observational study. J Antimicrob Chemother 67:22682273 [View Article][PubMed]
    [Google Scholar]
  5. Bialek R., Konrad F., Kern J., Aepinus C., Cecenas L., Gonzalez G. M., Just-Nübling G., Willinger B., Presterl E. et al. 2005; PCR based identification and discrimination of agents of mucormycosis and aspergillosis in paraffin wax embedded tissue. J Clin Pathol 58:11801184 [View Article][PubMed]
    [Google Scholar]
  6. Bitar D., Van Cauteren D., Lanternier F., Dannaoui E., Che D., Dromer F., Desenclos J. C., Lortholary O. 2009; Increasing incidence of zygomycosis (mucormycosis), France, 1997-2006. Emerg Infect Dis 15:13951401 [View Article][PubMed]
    [Google Scholar]
  7. Chamilos G., Lewis R. E., Kontoyiannis D. P. 2008; Delaying amphotericin B-based frontline therapy significantly increases mortality among patients with hematologic malignancy who have zygomycosis. Clin Infect Dis 47:503509 [View Article][PubMed]
    [Google Scholar]
  8. de Pauw B. E., Viscoli C. 2011; Managing invasive fungal infections: relying on clinical instincts or on a rational navigation system?. J Antimicrob Chemother 66:i55i58 [View Article][PubMed]
    [Google Scholar]
  9. De Pauw B., Walsh T. J., Donnelly J. P., Stevens D. A., Edwards J. E., Calandra T., Pappas P. G., Maertens J., Lortholary O. et al. 2008; Revised definitions of invasive fungal disease from the European Organization for Research and Treatment of Cancer/Invasive Fungal Infections Cooperative Group and the National Institute of Allergy and Infectious Diseases Mycoses Study Group (EORTC/MSG) Consensus Group. Clin Infect Dis 46:18131821 [View Article][PubMed]
    [Google Scholar]
  10. Hammond S. P., Baden L. R., Marty F. M. 2011; Mortality in hematologic malignancy and hematopoietic stem cell transplant patients with mucormycosis, 2001 to 2009. Antimicrob Agents Chemother 55:50185021 [View Article][PubMed]
    [Google Scholar]
  11. Ibrahim A. S., Spellberg B., Walsh T. J., Kontoyiannis D. P. 2012; Pathogenesis of mucormycosis. Clin Infect Dis 54:S16S22 [View Article][PubMed]
    [Google Scholar]
  12. Kasai M., Harrington S. M., Francesconi A., Petraitis V., Petraitiene R., Beveridge M. G., Knudsen T., Milanovich J., Cotton M. P. et al. 2008; Detection of a molecular biomarker for zygomycetes by quantitative PCR assays of plasma, bronchoalveolar lavage, and lung tissue in a rabbit model of experimental pulmonary zygomycosis. J Clin Microbiol 46:36903702 [View Article][PubMed]
    [Google Scholar]
  13. Kontoyiannis D. P., Lionakis M. S., Lewis R. E., Chamilos G., Healy M., Perego C., Safdar A., Kantarjian H., Champlin R. et al. 2005; Zygomycosis in a tertiary-care cancer center in the era of Aspergillus-active antifungal therapy: a case-control observational study of 27 recent cases. J Infect Dis 191:13501360 [View Article][PubMed]
    [Google Scholar]
  14. Kontoyiannis D. P., Wessel V. C., Bodey G. P., Rolston K. V. 2000; Zygomycosis in the 1990s in a tertiary-care cancer center. Clin Infect Dis 30:851856 [View Article][PubMed]
    [Google Scholar]
  15. Kwon-Chung K. J. 2012; Taxonomy of fungi causing mucormycosis and entomophthoramycosis (zygomycosis) and nomenclature of the disease: molecular mycologic perspectives. Clin Infect Dis 54:S8S15 [View Article][PubMed]
    [Google Scholar]
  16. Lackner M., Caramalho R., Lass-Flörl C. 2014; Laboratory diagnosis of mucormycosis: current status and future perspectives. Future Microbiol 9:683695 [View Article][PubMed]
    [Google Scholar]
  17. Lanternier F., Dannaoui E., Morizot G., Elie C., Garcia-Hermoso D., Huerre M., Bitar D., Dromer F., Lortholary O. French Mycosis Study Group 2012; A global analysis of mucormycosis in France: the RetroZygo Study (2005–2007). Clin Infect Dis 54:S35S43 [View Article][PubMed]
    [Google Scholar]
  18. Legrand M., Gits-Muselli M., Boutin L., Garcia-Hermoso D., Maurel V., Soussi S., Benyamina M., Ferry A., Chaussard M. et al. 2016; Detection of circulating Mucorales DNA in critically ill burn patients: preliminary report of a screening strategy for early diagnosis and treatment. Clin Infect Dis 63:13121317 [View Article][PubMed]
    [Google Scholar]
  19. Meletiadis J., Roilides E. 2013; Rare invasive fungal infections: epidemiology, diagnosis and management. Curr Fungal Infect Rep 7:351360 [View Article]
    [Google Scholar]
  20. Millon L., Larosa F., Lepiller Q., Legrand F., Rocchi S., Daguindau E., Scherer E., Bellanger A. P., Leroy J., Grenouillet F. 2013; Quantitative polymerase chain reaction detection of circulating DNA in serum for early diagnosis of mucormycosis in immunocompromised patients. Clin Infect Dis 56:e95e101 [View Article][PubMed]
    [Google Scholar]
  21. Millon L., Herbrecht R., Grenouillet F., Morio F., Alanio A., Letscher-Bru V., Cassaing S., Chouaki T., Kauffmann-Lacroix C. et al. 2015; Early diagnosis and monitoring of mucormycosis by detection of circulating DNA in serum: retrospective analysis of 44 cases collected through the French Surveillance Network of Invasive Fungal Infections (RESSIF). Clin Microbiol Infect 22:810.e1810.e8 [View Article]
    [Google Scholar]
  22. Petrikkos G., Skiada A., Lortholary O., Roilides E., Walsh T. J., Kontoyiannis D. P. 2012; Epidemiology and clinical manifestations of mucormycosis. Clin Infect Dis 54:S23S34 [View Article][PubMed]
    [Google Scholar]
  23. Rickerts V., Just-Nübling G., Konrad F., Kern J., Lambrecht E., Böhme A., Jacobi V., Bialek R. 2006; Diagnosis of invasive aspergillosis and mucormycosis in immunocompromised patients by seminested PCR assay of tissue samples. Eur J Clin Microbiol Infect Dis 25:813 [View Article][PubMed]
    [Google Scholar]
  24. Rogers B. B., Alpert L. C., Hine E. A., Buffone G. J. 1990; Analysis of DNA in fresh and fixed tissue by the polymerase chain reaction. Am J Pathol 136:541548[PubMed]
    [Google Scholar]
  25. Springer J., Schloßnagel H., Heinz W., Doedt T., Soeller R., Einsele H., Loeffler J. 2012; A novel extraction method combining plasma with a whole-blood fraction shows excellent sensitivity and reproducibility for patients at high risk for invasive aspergillosis. J Clin Microbiol 50:25852591 [View Article][PubMed]
    [Google Scholar]
  26. Springer J., Morton C. O., Perry M., Heinz W. J., Paholcsek M., Alzheimer M., Rogers T. R., Barnes R. A., Einsele H. et al. 2013; Multicenter comparison of serum and whole-blood specimens for detection of Aspergillus DNA in high-risk hematological patients. J Clin Microbiol 51:14451450 [View Article][PubMed]
    [Google Scholar]
  27. Springer J., Goldenberger D., Schmidt F., Weisser M., Wehrle-Wieland E., Einsele H., Frei R., Löffler J. 2016a; Development and application of two independent real-time PCR assays to detect clinically relevant Mucorales species. J Med Microbiol 65:227234 [View Article][PubMed]
    [Google Scholar]
  28. Springer J., Lackner M., Nachbaur D., Girschikofsky M., Risslegger B., Mutschlechner W., Fritz J., Heinz W. J., Einsele H. et al. 2016b; Prospective multicentre PCR-based Aspergillus DNA screening in high-risk patients with and without primary antifungal mould prophylaxis. Clin Microbiol Infect 22:8086 [View Article][PubMed]
    [Google Scholar]
  29. White P. L., Bretagne S., Klingspor L., Melchers W. J., McCulloch E., Schulz B., Finnstrom N., Mengoli C., Barnes R. A. et al. 2010; Aspergillus PCR: one step closer to standardization. J Clin Microbiol 48:12311240 [View Article][PubMed]
    [Google Scholar]
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