1887

Abstract

For diagnosing invasive aspergillosis (IA), an increasing clinical problem in immunocompromised patients, molecular tools are gaining in importance. Detection of DNA in blood samples was investigated by a nested PCR assay in a murine model of experimentally induced IA. , the detection threshold of the PCR assay was determined in blood and organ homogenates of mice. After intravenous injection of conidia on different days, growth of colonies was determined in cultures of blood and organs from immunocompetent and immunosuppressed mice and DNA was detected from blood samples by a nested PCR assay. The detection threshold of the PCR assay was as low as 1 c.f.u. ml. The assay proved to be more sensitive than cultures of blood, with sensitivity rates between 17.6 and 87.5 % depending on the fungal burden. In conclusion, the nested PCR assay is superior to cultural methods in detecting spp. in murine blood samples.

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2004-08-01
2019-11-21
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References

  1. Ascioglu, S., Rex, J. H., de Pauw, B. & 17 other authors ( 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.[CrossRef]
    [Google Scholar]
  2. Becker, M. J., de Marie, S., Willemse, D., Verbrugh, H. A. & Bakker-Woudenberg, I. A. ( 2000;). Quantitative galactomannan detection is superior to PCR in diagnosing and monitoring invasive pulmonary aspergillosis in an experimental rat model. J Clin Microbiol 38, 1434–1438.
    [Google Scholar]
  3. Bodey, G., Bueltmann, B., Duguid, W. & 7 other authors ( 1992;). Fungal infections in cancer patients: an international autopsy survey. Eur J Clin Microbiol Infect Dis 11, 99–109.[CrossRef]
    [Google Scholar]
  4. Buchheidt, D., Baust, C., Skladny, H., Ritter, J., Suedhoff, T., Baldus, M., Seifarth, W., Leib-Moesch, C. & Hehlmann, R. ( 2001;). Detection of Aspergillus species in blood and bronchoalveolar lavage samples from immunocompromised patients by means of 2-step polymerase chain reaction: clinical results. Clin Infect Dis 33, 428–435.[CrossRef]
    [Google Scholar]
  5. Denning, D. W. ( 1998;). Invasive aspergillosis. Clin Infect Dis 26, 781–803.[CrossRef]
    [Google Scholar]
  6. Denning, D. W., Kibbler, C. C. & Barnes, R. A. ( 2003;). British Society for Medical Mycology proposed standards of care for patients with invasive fungal infections. Lancet Infect Dis 3, 230–240.[CrossRef]
    [Google Scholar]
  7. Einsele, H., Hebart, H., Roller, G. & 8 other authors ( 1997;). Detection and identification of fungal pathogens in blood by using molecular probes. J Clin Microbiol 35, 1353–1360.
    [Google Scholar]
  8. Girmenia, C., Nucci, M. & Martino, P. ( 2001;). Clinical significance of Aspergillus fungaemia in patients with haematological malignancies and invasive aspergillosis. Br J Haematol 114, 93–98.[CrossRef]
    [Google Scholar]
  9. Groll, A. H., Shah, P. M., Mentzel, C., Schneider, M., Just-Nuebling, G. & Huebner, K. ( 1996;). Trends in the postmortem epidemiology of invasive fungal infections at a university hospital. J Infect 33, 23–32.[CrossRef]
    [Google Scholar]
  10. Hanazawa, R., Murayama, S. Y. & Yamaguchi, H. ( 2000;). In-situ detection of Aspergillus fumigatus. J Med Microbiol 49, 285–290.
    [Google Scholar]
  11. Kretschmar, M., Buchheidt, D., Hof, H. & Nichterlein, T. ( 2001;). Galactomannan enzyme immunoassay for monitoring systemic infection with Aspergillus fumigatus in mice. Diagn Microbiol Infect Dis 41, 107–112.[CrossRef]
    [Google Scholar]
  12. Latge, J. P. ( 2001;). The pathobiology of Aspergillus fumigatus. Trends Microbiol 9, 382–389.[CrossRef]
    [Google Scholar]
  13. Lin, S. J., Schranz, J. & Teutsch, S. M. ( 2001;). Aspergillosis case-fatality rate: systematic review of the literature. Clin Infect Dis 32, 358–366.[CrossRef]
    [Google Scholar]
  14. Loeffler, J., Kloepfer, K., Hebart, H. & 7 other authors ( 2002;). Polymerase chain reaction detection of Aspergillus DNA in experimental models of invasive aspergillosis. J Infect Dis 185, 1203–1206.[CrossRef]
    [Google Scholar]
  15. Maertens, J., Verhaegen, J., Lagrou, K., Van Eldere, J. & Boogaerts, M. ( 2001;). Screening for circulating galactomannan as a noninvasive diagnostic tool for invasive aspergillosis in prolonged neutropenic patients and stem cell transplantation recipients: a prospective validation. Blood 97, 1604–1610.[CrossRef]
    [Google Scholar]
  16. Makimura, K., Murayama, S. Y. & Yamaguchi, H. ( 1994;). Detection of a wide range of medically important fungi by the polymerase chain reaction. J Med Microbiol 40, 358–364.[CrossRef]
    [Google Scholar]
  17. Mori, K., Kawasaki, T. & Yamashina, I. ( 1983;). Identification of the mannan-binding protein from rat livers as a hepatocyte protein distinct from the mannan receptor on sinusoidal cells. Arch Biochem Biophys 222, 542–552.[CrossRef]
    [Google Scholar]
  18. Ruhnke, M. & Maschmeyer, G. ( 2002;). Management of mycoses in patients with hematologic disease and cancer – review of the literature. Eur J Med Res 7, 227–235.
    [Google Scholar]
  19. Skladny, H., Buchheidt, D., Baust, C., Krieg-Schneider, F., Seifarth, W., Leib-Mosch, C. & Hehlmann, R. ( 1999;). Specific detection of Aspergillus species in blood and bronchoalveolar lavage samples of immunocompromised patients by two-step PCR. J Clin Microbiol 37, 3865–3871.
    [Google Scholar]
  20. Verweij, P. E., Rijs, A. J., De Pauw, B. E., Horrevorts, A. M., Hoogkamp-Korstanje, J. A. & Meis, J. F. ( 1995;). Clinical evaluation and reproducibility of the Pastorex Aspergillus antigen latex agglutination test for diagnosing invasive aspergillosis. J Clin Pathol 48, 474–476.[CrossRef]
    [Google Scholar]
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