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Diagnosis of invasive aspergillosis and mucormycosis in immunocompromised patients by seminested PCR assay of tissue samples

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Abstract

Aspergillosis and mucormycosis are the most common mold infections in patients with hematological malignancies. Infections caused by species of the genus Aspergillus and the order Mucorales require different antifungal treatments depending on the in vitro susceptibility of the causative strain. Cultures from biopsy specimens frequently do not grow fungal pathogens, even from histopathologically proven cases of invasive fungal infection. Two seminested PCR assays were evaluated by amplifying DNA of zygomycetes and Aspergillus spp. from organ biopsies of 21 immunocompromised patients. The PCR assays correctly identified five cases of invasive aspergillosis and six cases of mucormycosis. They showed evidence of double mold infection in two cases. Both assays were negative in five negative controls and in two patients with yeast infections. Sequencing of the PCR products was in accordance with culture results in all culture-positive cases. In six patients without positive cultures but with positive histopathology, sequencing suggested a causative organism. Detection of fungal DNA from biopsy specimens allows rapid identification of the causative organism of invasive aspergillosis and mucormycosis. The use of these PCR assays may allow guided antifungal treatment in patients with invasive mold infections.

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References

  1. Fridkin S, Jarvis W (1996) Epidemiology of nosocomial fungal infections. Clin Microbiol Rev 9:499–511

    PubMed  CAS  Google Scholar 

  2. Groll A, Shah PM, Mentzel C et al (1996) Trends in postmortem epidemiology of invasive fungal infections at a university hospital. J Infect 33:23–32

    Article  PubMed  CAS  Google Scholar 

  3. Marr K, Carter R, Crippa F et al (2002) Epidemiology and outcome of mould infections in hematopoietic stem cell transplant recipients. Clin Infect Dis 34:909–917

    Article  PubMed  Google Scholar 

  4. Sun QN, Fothergill AW, McCarthy DI et al (2002) In vitro activities of posaconazole, itraconazole, voriconazole, amphotericin B, and fluconazole against 37 clinical isolates of zygomycetes. Antimicrob Agents Chemother 46:1581–1582

    Article  PubMed  CAS  Google Scholar 

  5. Ascioglu S, Rex J, de Pauw B et al (2002) Defining opportunistic invasive fungal infections in immunocompromised patients with cancer and hematopoietic stem cell transplants: an international consensus. Clin Infect Dis 34:7–14

    Article  PubMed  CAS  Google Scholar 

  6. Nosari A, Anghilieri M, Carrafiello G, et al (2003) Utility of percutaneous lung biopsy for diagnosing filamentous fungal infections in hematologic malignancies. Haematologica 88:1405–1409

    PubMed  Google Scholar 

  7. Denning D (1998) Invasive aspergillosis. Clin Infect Dis 26:781–805

    Article  PubMed  CAS  Google Scholar 

  8. Ribes J, Vanover-Sams C, Baker D (2000) Zygomycetes in human disease. Clin Microbiol Rev 13:236–301

    Article  PubMed  CAS  Google Scholar 

  9. Bialek R, Fischer J, Feucht A et al (2001) Diagnosis and monitoring of murine histoplasmosis by a nested PCR assay. J Clin Microbiol 29:1506–1509

    Article  Google Scholar 

  10. Loeffler J, Kloepfer K, Hebart H et al (2001) Polymerase chain reaction detection of Aspergillus DNA in experimental models of invasive aspergillosis. J Infect Dis 185:1203–1206

    Article  Google Scholar 

  11. Bialek R, Konrad F, Kern J et al (2005) PCR-based identification and discrimination of mucormycosis and aspergillosis in paraffin-embedded tissue. J Clin Pathol 58:1180–1184

    Article  PubMed  CAS  Google Scholar 

  12. Bretagne S, Costa J, Marmorat-Khuong A et al (1995) Detection of Aspergillus species DNA in bronchoalveolar lavage samples by competitive PCR. J Clin Microbiol 33:1164–1168

    PubMed  CAS  Google Scholar 

  13. Martino R, Subriá M, Rovira M et al (2002) Invasive fungal infections after allogeneic peripheral blood stem cell transplantation: incidence and risk factors in 395 patients. Br J Haematol 116:475–482

    Article  PubMed  Google Scholar 

  14. Herbrecht R, Denning D, Patterson T et al (2002) Voriconazole versus amphotericin B for primary therapy of invasive aspergillosis. N Engl J Med 347:408–415

    Article  PubMed  CAS  Google Scholar 

  15. Dannaoui E, Borel E, Persat F et al (2000) Amphotericin B resistance of Aspergillus terreus in a murine model of disseminated aspergillosis. J Med Microbiol 49:601–606

    PubMed  CAS  Google Scholar 

  16. Iwen P, Rupp M, Langnas A et al (2003) Invasive pulmonary aspergillosis due to Aspergillus terreus: 12-year experience and review of the literature. Clin Infect Dis 26:1092–1097

    Article  Google Scholar 

  17. Kim K, Lee M, Kim J et al (2002) Importance of open lung biopsy in the diagnosis of invasive pulmonary aspergillosis in patients with hematologic malignancies. Am J Hematol 71:75–79

    Article  PubMed  Google Scholar 

  18. Pidhorecky I, Urschel J, Anderson T (2003) Resection of invasive pulmonary aspergillosis in immunocompromised patients. Ann Surg Oncol 7:312–317

    Article  Google Scholar 

  19. Yamazaki T, Kume H, Murase S et al (1999) Epidemiology of visceral mycoses: analysis of data in annual of the pathological autopsy cases in Japan. J Clin Microbiol 37:1732–1738

    PubMed  CAS  Google Scholar 

  20. Gómez-López A, Cuenca-Estrella M, Monzón A et al (2001) In vitro susceptibility of clinical isolates of Zygomycota to amphotericin B, flucytosine, itraconazole and voriconazole. J Antimicrob Chemother 48:919–921

    Article  PubMed  Google Scholar 

  21. Pfaller M, Marco F, Messer S et al (1998) In vitro activity of two echinocandin derivatives, LY303366 and MK-0991 (L-743,792), against clinical isolates of Aspergillus, Fusarium, Rhizopus and other filamentous fungi. Diagn Microbiol Infect Dis 30:251–255

    Article  PubMed  CAS  Google Scholar 

  22. Marty FM, Cosimi LA, Baden LR (2004) Breakthrough zygomycosis after voriconazole treatment in recipients of hematopoietic stem-cell transplants. New Engl J Med 26:950–952

    Article  Google Scholar 

  23. Rickerts V, Böhme A, Viertel A et al (2000) Cluster of pulmonary infections caused by Cunninghamella bertholletiae in immunocompromised patients. Clin Infect Dis 31:910–913

    Article  PubMed  CAS  Google Scholar 

  24. Rickerts V, Loeffler J, Böhme A et al (2001) Diagnosis of disseminated zygomycosis using a polymerase chain reaction assay. Eur J Clin Microbiol Infect Dis 20:744–745

    Article  PubMed  CAS  Google Scholar 

  25. Herbrecht R, Letscher-Bru R, Bowden R et al (2001) Treatment of 21 cases of invasive mucormycosis with amphotericin B colloidal dispersion. Eur J Clin Microbiol Infect Dis 20:460–466

    Article  PubMed  CAS  Google Scholar 

  26. Dannaoui E, Meletiadis J, Mouton J et al (2003) In vitro susceptibilities of zygomycetes to conventional and new antifungals. J Antimicrob Chemother 51:45–52

    Article  PubMed  CAS  Google Scholar 

  27. Kontoyiannis D, Wessel W, Bodey G et al (2000) Zygomycosis in the 1990s in a tertiary-care cancer center. Clin Infect Dis 30:851–856

    Article  PubMed  CAS  Google Scholar 

  28. Imhof A, Schaer C, Schoedon G et al (2003) Rapid detection of pathogenic fungi from clinical specimens using LightCycler Real Time Fluorescence PCR. Eur J Clin Microbiol Infect Dis 22:558–560

    Article  PubMed  CAS  Google Scholar 

  29. Willinger B, Obradovic A, Selitsch B et al (2003) Detection and identification of fungi from fungus balls of the maxillary sinus by molecular techniques. J Clin Microbiol 41:581–585

    Article  PubMed  CAS  Google Scholar 

  30. Bialek R, Gonzalez GM, Begerow D, Zelck UE (2005) Coccidioidomycosis and blastomycosis: advances in molecular diagnosis (related articles, links). FEMS Immunol Med Microbiol 45:355–360

    Article  PubMed  CAS  Google Scholar 

  31. Bialek R, Weiss M, Bekure-Nemariam K et al (2002) Detection of Cryptococcus neoformans DNA in tissue samples by nested and real-time PCR assays. Clin Diagn Lab Immunol 9:461–469

    Article  PubMed  CAS  Google Scholar 

  32. Hayden RT, Qian X, Procop GW et al (2002) In situ hybridization for the identification of filamentous fungi in tissue section. Diagn Mol Pathol 11:119–126

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgement

The data presented here are part of the doctoral thesis of Franziska Konrad.

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Authors and Affiliations

  1. Med.Klinik II/Infektiologie, Klinikum der J.W. Goethe Universität, Theodor Stern Kai 7, 60590, Frankfurt, Germany

    V. Rickerts & G. Just-Nübling

  2. Institut für Tropenmedizin, Universitätsklinikum Tübingen, Keplerstrasse 15, 72074, Tübingen, Germany

    F. Konrad, J. Kern, V. Jacobi & R. Bialek

  3. Institut für Pathologie, Klinikum der J.W. Goethe Universität Frankfurt, Theodor Stern Kai 7, 60590, Frankfurt, Germany

    E. Lambrecht

  4. Med. Klinik II/Hämatologie, Klinikum der J.W. Goethe Universität, Theodor Stern Kai 7, 60590, Frankfurt, Germany

    A. Böhme

  5. Institut für Radiologie, Klinikum der J.W. Goethe Universität, Theodor Stern Kai 7, 60590, Frankfurt, Germany

    R. Bialek

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  1. V. Rickerts
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  2. G. Just-Nübling
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  3. F. Konrad
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  4. J. Kern
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  6. A. Böhme
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  7. V. Jacobi
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  8. R. Bialek
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Correspondence to V. Rickerts.

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Rickerts, V., Just-Nübling, G., Konrad, F. et al. Diagnosis of invasive aspergillosis and mucormycosis in immunocompromised patients by seminested PCR assay of tissue samples. Eur J Clin Microbiol Infect Dis 25, 8–13 (2006). https://doi.org/10.1007/s10096-005-0078-7

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