This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Skladny, H.
Right arrow Articles by Hehlmann, R.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Skladny, H.
Right arrow Articles by Hehlmann, R.

 Previous Article  |  Next Article 

Journal of Clinical Microbiology, December 1999, p. 3865-3871, Vol. 37, No. 12
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Specific Detection of Aspergillus Species in Blood and Bronchoalveolar Lavage Samples of Immunocompromised Patients by Two-Step PCR

Heyko Skladny,1 Dieter Buchheidt,1,* Corinna Baust,1 Frank Krieg-Schneider,1,dagger Wolfgang Seifarth,1 Christine Leib-Mösch,1,2 and Rüdiger Hehlmann1

III. Medizinische Klink, Klinikum Mannheim, University of Heidelberg, D-68305 Mannheim,1 and Institute of Molecular Virology, GSF-National Research Center for Environment and Health, D-85764 Neuherberg,2 Germany

Received 22 February 1999/Returned for modification 12 July 1999/Accepted 9 August 1999

The increasing incidence of aspergillosis, a life-threatening infection in immunocompromised patients, emphasizes the need to improve the currently limited diagnostic tools. We developed a two-step PCR assay that specifically amplifies a region of the 18S rRNA gene that is highly conserved in Aspergillus species. A number of primers with the least homology to equivalent human or Candida gene sequences were screened for the pairs that gave the highest sensitivity and specificity. No cross-reaction with the wide range of fungal and bacterial pathogens so far tested was observed. This assay allows direct and rapid detection of down to 10 fg of Aspergillus DNA corresponding to 1 to 5 CFU per ml of blood. A total of 315 blood and bronchoalveolar lavage samples from 140 subjects, including 93 patients at risk for invasive fungal disease, were screened. The result was a 100% correlation between positive histology, culture, or high-resolution computed tomography findings and PCR results. The test specificity was 89%. Our data point to the considerable potential clinical value of this simple, specific, rapid, and inexpensive PCR assay for improving the means of early diagnosis of systemic aspergillosis in high-risk patients.

* Corresponding author. Mailing address: III. Medizinische Klinik, Klinikum Mannheim der Universität Heidelberg, Wiesbadenerstraße 7-11, D-68305 Mannheim, Germany. Phone: 49-621-383-4115. Fax: 49-621-383-4201. E-mail: dieter.buchheidt{at}urz.uni-heidelberg.de.

dagger Present address: QIAGEN GmbH, Hilden, Germany.

Journal of Clinical Microbiology, December 1999, p. 3865-3871, Vol. 37, No. 12
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Spiess, B., Seifarth, W., Hummel, M., Frank, O., Fabarius, A., Zheng, C., Morz, H., Hehlmann, R., Buchheidt, D. (2007). DNA Microarray-Based Detection and Identification of Fungal Pathogens in Clinical Samples from Neutropenic Patients. J. Clin. Microbiol. 45: 3743-3753 [Abstract] [Full Text]  
  • Hummel, M., Spiess, B., Kentouche, K., Niggemann, S., Bohm, C., Reuter, S., Kiehl, M., Morz, H., Hehlmann, R., Buchheidt, D. (2006). Detection of Aspergillus DNA in Cerebrospinal Fluid from Patients with Cerebral Aspergillosis by a Nested PCR Assay. J. Clin. Microbiol. 44: 3989-3993 [Abstract] [Full Text]  
  • Huang, A., Li, J.-W., Shen, Z.-Q., Wang, X.-W., Jin, M. (2006). High-throughput identification of clinical pathogenic fungi by hybridization to an oligonucleotide microarray.. J. Clin. Microbiol. 44: 3299-3305 [Abstract] [Full Text]  
  • Playford, E. G., Kong, F., Sun, Y., Wang, H., Halliday, C., Sorrell, T. C. (2006). Simultaneous detection and identification of Candida, Aspergillus, and cryptococcus species by reverse line blot hybridization.. J. Clin. Microbiol. 44: 876-880 [Abstract] [Full Text]  
  • Kasai, M., Francesconi, A., Petraitiene, R., Petraitis, V., Kelaher, A. M., Kim, H.-s., Meletiadis, J., Sein, T., Bacher, J., Walsh, T. J. (2006). Use of Quantitative Real-Time PCR To Study the Kinetics of Extracellular DNA Released from Candida albicans, with Implications for Diagnosis of Invasive Candidiasis. J. Clin. Microbiol. 44: 143-150 [Abstract] [Full Text]  
  • Scotter, J. M., Chambers, S. T. (2005). Comparison of Galactomannan Detection, PCR-Enzyme-Linked Immunosorbent Assay, and Real-Time PCR for Diagnosis of Invasive Aspergillosis in a Neutropenic Rat Model and Effect of Caspofungin Acetate. CVI 12: 1322-1327 [Abstract] [Full Text]  
  • Halliday, C., Wu, Q. X., James, G., Sorrell, T. (2005). Development of a Nested Qualitative Real-Time PCR Assay To Detect Aspergillus Species DNA in Clinical Specimens. J. Clin. Microbiol. 43: 5366-5368 [Abstract] [Full Text]  
  • Hsiao, C. R., Huang, L., Bouchara, J.-P., Barton, R., Li, H. C., Chang, T. C. (2005). Identification of Medically Important Molds by an Oligonucleotide Array. J. Clin. Microbiol. 43: 3760-3768 [Abstract] [Full Text]  
  • White, P. L., Archer, A. E., Barnes, R. A. (2005). Comparison of Non-Culture-Based Methods for Detection of Systemic Fungal Infections, with an Emphasis on Invasive Candida Infections. J. Clin. Microbiol. 43: 2181-2187 [Abstract] [Full Text]  
  • Bu, R., Sathiapalan, R. K, Ibrahim, M. M, Al-Mohsen, I., Almodavar, E., Gutierrez, M. I, Bhatia, K. (2005). Monochrome LightCycler PCR assay for detection and quantification of five common species of Candida and Aspergillus. J Med Microbiol 54: 243-248 [Abstract] [Full Text]  
  • Polzehl, D., Weschta, M., Podbielski, A., Riechelmann, H., Rimek, D. (2005). Fungus culture and PCR in nasal lavage samples of patients with chronic rhinosinusitis. J Med Microbiol 54: 31-37 [Abstract] [Full Text]  
  • Lass-Florl, C., Gunsilius, E., Gastl, G., Bonatti, H., Freund, M. C., Gschwendtner, A., Kropshofer, G., Dierich, M. P., Petzer, A. (2004). Diagnosing Invasive Aspergillosis during Antifungal Therapy by PCR Analysis of Blood Samples. J. Clin. Microbiol. 42: 4154-4157 [Abstract] [Full Text]  
  • de Aguirre, L., Hurst, S. F., Choi, J. S., Shin, J. H., Hinrikson, H. P., Morrison, C. J. (2004). Rapid Differentiation of Aspergillus Species from Other Medically Important Opportunistic Molds and Yeasts by PCR-Enzyme Immunoassay. J. Clin. Microbiol. 42: 3495-3504 [Abstract] [Full Text]  
  • Hummel, M., Baust, C., Kretschmar, M., Nichterlein, T., Schleiermacher, D., Spiess, B., Skladny, H., Morz, H., Hehlmann, R., Buchheidt, D. (2004). Detection of Aspergillus DNA by a nested PCR assay is superior to blood culture in an experimental murine model of invasive aspergillosis. J Med Microbiol 53: 803-806 [Abstract] [Full Text]  
  • Challier, S., Boyer, S., Abachin, E., Berche, P. (2004). Development of a Serum-Based Taqman Real-Time PCR Assay for Diagnosis of Invasive Aspergillosis. J. Clin. Microbiol. 42: 844-846 [Abstract] [Full Text]  
  • O'Sullivan, C. E., Kasai, M., Francesconi, A., Petraitis, V., Petraitiene, R., Kelaher, A. M., Sarafandi, A. A., Walsh, T. J. (2003). Development and Validation of a Quantitative Real-Time PCR Assay Using Fluorescence Resonance Energy Transfer Technology for Detection of Aspergillus fumigatus in Experimental Invasive Pulmonary Aspergillosis. J. Clin. Microbiol. 41: 5676-5682 [Abstract] [Full Text]  
  • Rantakokko-Jalava, K., Laaksonen, S., Issakainen, J., Vauras, J., Nikoskelainen, J., Viljanen, M. K., Salonen, J. (2003). Semiquantitative Detection by Real-Time PCR of Aspergillus fumigatus in Bronchoalveolar Lavage Fluids and Tissue Biopsy Specimens from Patients with Invasive Aspergillosis. J. Clin. Microbiol. 41: 4304-4311 [Abstract] [Full Text]  
  • Sanguinetti, M., Posteraro, B., Pagano, L., Pagliari, G., Fianchi, L., Mele, L., La Sorda, M., Franco, A., Fadda, G. (2003). Comparison of Real-Time PCR, Conventional PCR, and Galactomannan Antigen Detection by Enzyme-Linked Immunosorbent Assay Using Bronchoalveolar Lavage Fluid Samples from Hematology Patients for Diagnosis of Invasive Pulmonary Aspergillosis. J. Clin. Microbiol. 41: 3922-3925 [Abstract] [Full Text]  
  • Spiess, B., Buchheidt, D., Baust, C., Skladny, H., Seifarth, W., Zeilfelder, U., Leib-Mosch, C., Morz, H., Hehlmann, R. (2003). Development of a LightCycler PCR Assay for Detection and Quantification of Aspergillus fumigatus DNA in Clinical Samples from Neutropenic Patients. J. Clin. Microbiol. 41: 1811-1818 [Abstract] [Full Text]  
  • Luo, G., Mitchell, T. G. (2002). Rapid Identification of Pathogenic Fungi Directly from Cultures by Using Multiplex PCR. J. Clin. Microbiol. 40: 2860-2865 [Abstract] [Full Text]  
  • Yeo, S. F., Wong, B. (2002). Current Status of Nonculture Methods for Diagnosis of Invasive Fungal Infections. Clin. Microbiol. Rev. 15: 465-484 [Abstract] [Full Text]  
  • Raad, I., Hanna, H., Huaringa, A., Sumoza, D., Hachem, R., Albitar, M. (2002). Diagnosis of Invasive Pulmonary Aspergillosis Using Polymerase Chain Reaction-Based Detection of Aspergillus in BAL*. Chest 121: 1171-1176 [Abstract] [Full Text]  
  • Chang, H. C., Leaw, S. N., Huang, A. H., Wu, T. L., Chang, T. C. (2001). Rapid Identification of Yeasts in Positive Blood Cultures by a Multiplex PCR Method. J. Clin. Microbiol. 39: 3466-3471 [Abstract] [Full Text]  
  • Hayette, M.-P., Vaira, D., Susin, F., Boland, P., Christiaens, G., Melin, P., De Mol, P. (2001). Detection of Aspergillus Species DNA by PCR in Bronchoalveolar Lavage Fluid. J. Clin. Microbiol. 39: 2338-2340 [Abstract] [Full Text]  
  • Reichenberger, F., Habicht, J.M., Gratwohl, A., Tamm, M. (2001). Diagnosis and treatment of invasive pulmonary aspergillosis in neutropenic patients. Eur Respir J 19: 743-755 [Abstract] [Full Text]  
  • Park, S., Wong, M., Marras, S. A. E., Cross, E. W., Kiehn, T. E., Chaturvedi, V., Tyagi, S., Perlin, D. S. (2000). Rapid Identification of Candida dubliniensis Using a Species-Specific Molecular Beacon. J. Clin. Microbiol. 38: 2829-2836 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»