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Clinical performance of a commercial real-time PCR assay for Aspergillus DNA detection in serum samples from high-risk patients: comparison with a galactomannan enzyme immunoassay

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Abstract

We investigated the clinical performance of a polymerase chain reaction (PCR)-based commercial platform, the Myconostica MycAssay™ Aspergillus (MAP), for fungal DNA detection in the serum of patients at risk of invasive aspergillosis (IA). Sixty-four hospitalized patients were prospectively enrolled and a total of 71 different episodes were investigated (30 episodes were clinically/microbiologically classified as IA and 41 as control episodes). When MAP was compared to the galactomannan (GM) assay, no significant differences were found in terms of sensitivity (46.7 % vs. 50.0 %), specificity (97.6 % vs. 95.1 %), positive predictive value (PPV) (93.3 % vs. 88.2 %), and negative predictive value (NPV) (71.4 % vs. 72.2 %). The corresponding areas under the curve (AUC) of the receiver operating characteristic (ROC) curves were also superimposable. Overall, because of the good agreement between the two assays and considering the high specificity and PPV of the MAP, we suggest the use of this PCR-based platform as a second-level examination for the evaluation of clinically undefined cases where culture or GM have provided positive results.

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References

  1. Vincent J-L, Rello J, Marshall J et al (2009) International study of the prevalence and outcomes of infection in intensive care units. JAMA 302:2323–2329. doi:10.1001/jama.2009.1754

    Article  CAS  PubMed  Google Scholar 

  2. Baddley JW, Andes DR, Marr KA et al (2010) Factors associated with mortality in transplant patients with invasive aspergillosis. Clin Infect Dis 50:1559–1567. doi:10.1086/652768

    Article  PubMed  PubMed Central  Google Scholar 

  3. Tortorano AM, Dho G, Prigitano A et al (2012) Invasive fungal infections in the intensive care unit: a multicentre, prospective, observational study in Italy (2006–2008). Mycoses 55:73–79. doi:10.1111/j.1439-0507.2011.02044.x

    Article  PubMed  Google Scholar 

  4. Montagna MT, De Giglio O, Napoli C et al (2012) Invasive fungal infections in patients with hematologic malignancies (Aurora Project): lights and shadows during 18-months surveillance. Int J Mol Sci 13:774–787. doi:10.3390/ijms13010774

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. von Eiff M, Roos N, Schulten R, Hesse M, Zühlsdorf M, van de Loo J (1995) Pulmonary aspergillosis: early diagnosis improves survival. Respiration 62:341–347

    Article  Google Scholar 

  6. Leeflang MM, Debets-Ossenkopp YJ, Visser CE et al (2008) Galactomannan detection for invasive aspergillosis in immunocompromized patients. Cochrane Database Syst Rev (4):CD007394. doi:10.1002/14651858.CD007394

  7. Zou M, Tang L, Zhao S et al (2012) Systematic review and meta-analysis of detecting galactomannan in bronchoalveolar lavage fluid for diagnosing invasive aspergillosis. PLoS One 7:e43347. doi:10.1371/journal.pone.0043347

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Ostrosky-Zeichner L, Alexander BD, Kett DH et al (2005) Multicenter clinical evaluation of the (1–>3) beta-D-glucan assay as an aid to diagnosis of fungal infections in humans. Clin Infect Dis 41:654–659. doi:10.1086/432470

    Article  CAS  PubMed  Google Scholar 

  9. Onishi A, Sugiyama D, Kogata Y et al (2012) Diagnostic accuracy of serum 1,3-β-D-glucan for Pneumocystis jiroveci pneumonia, invasive candidiasis, and invasive aspergillosis: systematic review and meta-analysis. J Clin Microbiol 50:7–15. doi:10.1128/JCM.05267-11

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Mengoli C, Cruciani M, Barnes RA, Loeffler J, Donnelly JP (2009) Use of PCR for diagnosis of invasive aspergillosis: systematic review and meta-analysis. Lancet Infect Dis 9:89–96. doi:10.1016/S1473-3099(09)70019-2

    Article  CAS  PubMed  Google Scholar 

  11. Maertens J, Marchetti O, Herbrecht R et al (2011) European guidelines for antifungal management in leukemia and hematopoietic stem cell transplant recipients: summary of the ECIL 3—2009 update. Bone Marrow Transplant 46:709–718. doi:10.1038/bmt.2010.175

    Article  CAS  PubMed  Google Scholar 

  12. Dellinger RP, Levy MM, Rhodes A et al (2013) Surviving sepsis campaign: international guidelines for management of severe sepsis and septic shock: 2012. Crit Care Med 41:580–637. doi:10.1097/CCM.0b013e31827e83af

    Article  PubMed  Google Scholar 

  13. De Pauw B, Walsh TJ, Donnelly JP 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 46:1813–1821. doi:10.1086/588660

    Article  PubMed  PubMed Central  Google Scholar 

  14. Meersseman W, Vandecasteele SJ, Wilmer A, Verbeken E, Peetermans WE, Van Wijngaerden E (2004) Invasive aspergillosis in critically ill patients without malignancy. Am J Respir Crit Care Med 170:621–625. doi:10.1164/rccm.200401-093OC

    Article  PubMed  Google Scholar 

  15. Meersseman W, Lagrou K, Maertens J, Van Wijngaerden E (2007) Invasive aspergillosis in the intensive care unit. Clin Infect Dis 45:205–216. doi:10.1086/518852

    Article  PubMed  Google Scholar 

  16. Torelli R, Sanguinetti M, Moody A et al (2011) Diagnosis of invasive aspergillosis by a commercial real-time PCR assay for Aspergillus DNA in bronchoalveolar lavage fluid samples from high-risk patients compared to a galactomannan enzyme immunoassay. J Clin Microbiol 49:4273–4278. doi:10.1128/JCM.05026-11

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. White PL, Perry MD, Moody A, Follett SA, Morgan G, Barnes RA (2011) Evaluation of analytical and preliminary clinical performance of Myconostica MycAssay Aspergillus when testing serum specimens for diagnosis of invasive aspergillosis. J Clin Microbiol 49:2169–2174. doi:10.1128/JCM.00101-11

    Article  PubMed  PubMed Central  Google Scholar 

  18. Kedzierska A, Kochan P, Pietrzyk A, Kedzierska J (2007) Current status of fungal cell wall components in the immunodiagnostics of invasive fungal infections in humans: galactomannan, mannan and (1–>3)-beta-D-glucan antigens. Eur J Clin Microbiol Infect Dis 26:755–766. doi:10.1007/s10096-007-0373-6

    Article  CAS  PubMed  Google Scholar 

  19. Marr KA, Laverdiere M, Gugel A, Leisenring W (2005) Antifungal therapy decreases sensitivity of the Aspergillus galactomannan enzyme immunoassay. Clin Infect Dis 40:1762–1769. doi:10.1086/429921

    Article  CAS  PubMed  Google Scholar 

  20. Lass-Flörl C, Gunsilius E, Gastl G et al (2004) Diagnosing invasive aspergillosis during antifungal therapy by PCR analysis of blood samples. J Clin Microbiol 42:4154–4157. doi:10.1128/JCM.42.9.4154-4157.2004

    Article  PubMed  PubMed Central  Google Scholar 

  21. White PL, Mengoli C, Bretagne S et al (2011) Evaluation of Aspergillus PCR protocols for testing serum specimens. J Clin Microbiol 49:3842–3848. doi:10.1128/JCM.05316-11

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Rogers TR, Morton CO, Springer J et al (2013) Combined real-time PCR and galactomannan surveillance improves diagnosis of invasive aspergillosis in high risk patients with haematological malignancies. Br J Haematol 161:517–524. doi:10.1111/bjh.12285

    Article  CAS  PubMed  Google Scholar 

  23. White PL, Perry MD, Loeffler J et al (2010) Critical stages of extracting DNA from Aspergillus fumigatus in whole-blood specimens. J Clin Microbiol 48:3753–3755. doi:10.1128/JCM.01466-10

    Article  PubMed  PubMed Central  Google Scholar 

  24. Orsi CF, Gennari W, Venturelli C et al (2012) Performance of 2 commercial real-time polymerase chain reaction assays for the detection of Aspergillus and Pneumocystis DNA in bronchoalveolar lavage fluid samples from critical care patients. Diagn Microbiol Infect Dis 73:138–143. doi:10.1016/j.diagmicrobio.2012.03.001

    Article  CAS  PubMed  Google Scholar 

  25. Guinea J, Padilla C, Escribano P et al (2013) Evaluation of MycAssay™ Aspergillus for diagnosis of invasive pulmonary aspergillosis in patients without hematological cancer. PLoS One 8:e61545. doi:10.1371/journal.pone.0061545

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  26. Hidalgo A, Parody R, Martino R et al (2009) Correlation between high-resolution computed tomography and galactomannan antigenemia in adult hematologic patients at risk for invasive aspergillosis. Eur J Radiol 71:55–60. doi:10.1016/j.ejrad.2008.03.026

    Article  CAS  PubMed  Google Scholar 

  27. Yamakami Y, Hashimoto A, Yamagata E et al (1998) Evaluation of PCR for detection of DNA specific for Aspergillus species in sera of patients with various forms of pulmonary aspergillosis. J Clin Microbiol 36:3619–3623

    CAS  PubMed  PubMed Central  Google Scholar 

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Acknowledgments

We thank Dr. Andrea Ardizzoni for the critical reading of the manuscript and suggestions.

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The authors declare that they have no conflict of interest.

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Correspondence to E. Blasi.

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Pini, P., Bettua, C., Orsi, C.F. et al. Clinical performance of a commercial real-time PCR assay for Aspergillus DNA detection in serum samples from high-risk patients: comparison with a galactomannan enzyme immunoassay. Eur J Clin Microbiol Infect Dis 34, 131–136 (2015). https://doi.org/10.1007/s10096-014-2211-y

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  • DOI: https://doi.org/10.1007/s10096-014-2211-y

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