This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Bretagne, S. Articles by Fleury-Feith, J. Search for Related Content PubMed PubMed Citation Articles by Bretagne, S. Articles by Fleury-Feith, J.

 Previous Article  |  Next Article 

Journal of Clinical Microbiology, 05 1995, 1164-1168, Vol 33, No. 5
Copyright © 1995 by the American Society for Microbiology. All rights reserved.

Detection of Aspergillus species DNA in bronchoalveolar lavage samples by competitive PCR

S Bretagne, JM Costa, A Marmorat-Khuong, F Poron, C Cordonnier, M Vidaud and J Fleury-Feith
Laboratoire de Parasitologie-Mycologie, Hopital Henri Mondor, Creteil, France.

A competitive PCR assay involving the use of bronchoalveolar lavage (BAL) samples for the diagnosis of invasive pulmonary aspergillosis (IPA) was developed. For this purpose, a 1-kb mitochondrial DNA fragment of Aspergillus fumigatus was sequenced. The primers used allowed amplification of A. fumigatus, A. flavus, A. terreus, and A. niger DNAs but not DNAs of other fungi and yeasts. BAL samples from 55 consecutively enrolled patients were tested. Three samples were excluded because of failure of correct amplification of the internal competitive control. Of 28 immunocompromised patients, 6 were PCR positive; 3 died of IPA and their BAL cultures yielded A. fumigatus; and 3 were culture negative and did not develop IPA. Of 15 human immunodeficiency virus-positive patients and 9 immunocompetent patients, 5 and 4, respectively, were both PCR positive and culture negative, and none developed aspergillosis. Thus, PCR confirmed IPA in three patients but gave positive results for 25% (12 of 49) of the patients who did not develop aspergillosis. The predictive value of PCR- positive results seems low for patients at risk for aspergillosis. Moreover, the risk of contamination of reaction buffers or biological samples with Aspergillus conidia seems high and has to be weighed in regard to the potential diagnostic benefit of PCR testing as a routine procedure.

This article has been cited by other articles:

• Bulpa, P., Dive, A., Sibille, Y. (2007). Invasive pulmonary aspergillosis in patients with chronic obstructive pulmonary disease. Eur Respir J 30: 782-800 [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]  
• Bialek, R, Konrad, F, Kern, J, Aepinus, C, Cecenas, L, Gonzalez, G M, Just-Nubling, G, Willinger, B, Presterl, E, Lass-Florl, C, Rickerts, V (2005). PCR based identification and discrimination of agents of mucormycosis and aspergillosis in paraffin wax embedded tissue. J. Clin. Pathol. 58: 1180-1184 [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]  
• 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]  
• 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]  
• 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]  
• Costa, C., Costa, J.-M., Desterke, C., Botterel, F., Cordonnier, C., Bretagne, S. (2002). Real-Time PCR Coupled with Automated DNA Extraction and Detection of Galactomannan Antigen in Serum by Enzyme-Linked Immunosorbent Assay for Diagnosis of Invasive Aspergillosis. J. Clin. Microbiol. 40: 2224-2227 [Abstract] [Full Text]  
• Soubani, A. O., Chandrasekar, P. H. (2002). The Clinical Spectrum of Pulmonary Aspergillosis*. Chest 121: 1988-1999 [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]  
• Bowman, J. C., Abruzzo, G. K., Anderson, J. W., Flattery, A. M., Gill, C. J., Pikounis, V. B., Schmatz, D. M., Liberator, P. A., Douglas, C. M. (2001). Quantitative PCR Assay To Measure Aspergillus fumigatus Burden in a Murine Model of Disseminated Aspergillosis: Demonstration of Efficacy of Caspofungin Acetate. Antimicrob. Agents Chemother. 45: 3474-3481 [Abstract] [Full Text]  
• Zhao, J., Kong, F., Li, R., Wang, X., Wan, Z., Wang, D. (2001). Identification of Aspergillus fumigatus and Related Species by Nested PCR Targeting Ribosomal DNA Internal Transcribed Spacer Regions. J. Clin. Microbiol. 39: 2261-2266 [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]  
• Verweij, P. E., Weemaes, C. M., Curfs, J. H. A. J., Bretagne, S., Meis, J. F. G. M. (2000). Failure To Detect Circulating Aspergillus Markers in a Patient with Chronic Granulomatous Disease and Invasive Aspergillosis. J. Clin. Microbiol. 38: 3900-3901 [Abstract] [Full Text]  
• Henry, T., Iwen, P. C., Hinrichs, S. H. (2000). Identification of Aspergillus Species Using Internal Transcribed Spacer Regions 1 and 2. J. Clin. Microbiol. 38: 1510-1515 [Abstract] [Full Text]  
• Skladny, H., Buchheidt, D., Baust, C., Krieg-Schneider, F., Seifarth, W., Leib-Mösch, 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 [Abstract] [Full Text]  
• Hue, F.-X., Huerre, M., Rouffault, M. A., de Bievre, C. (1999). Specific Detection of Fusarium Species in Blood and Tissues by a PCR Technique. J. Clin. Microbiol. 37: 2434-2438 [Abstract] [Full Text]  
• Latge, J.-P. (1999). Aspergillus fumigatus and Aspergillosis. Clin. Microbiol. Rev. 12: 310-350 [Abstract] [Full Text]  
• Kawamura, S., Maesaki, S., Noda, T., Hirakata, Y., Tomono, K., Tashiro, T., Kohno, S. (1999). Comparison between PCR and Detection of Antigen in Sera for Diagnosis of Pulmonary Aspergillosis. J. Clin. Microbiol. 37: 218-220 [Abstract] [Full Text]  
• Van Burik, J.-A., Myerson, D., Schreckhise, R. W., Bowden, R. A. (1998). Panfungal PCR Assay for Detection of Fungal Infection in Human Blood Specimens. J. Clin. Microbiol. 36: 1169-1175 [Abstract] [Full Text]