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 Costa, C.
Right arrow Articles by Bretagne, S.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Costa, C.
Right arrow Articles by Bretagne, S.

 Previous Article  |  Next Article 

Journal of Clinical Microbiology, June 2002, p. 2224-2227, Vol. 40, No. 6
0095-1137/02/$04.00+0     DOI: 10.1128/JCM.40.6.2224-2227.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

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

Catherine Costa,1 Jean-Marc Costa,1,2 Christophe Desterke,1 Françoise Botterel,1 Catherine Cordonnier,3 and Stéphane Bretagne1*

Laboratoire de Parasitologie-Mycologie and UMR BIPAR 956,1 Service d'Hématologie Clinique, Hôpital H. Mondor-APHP, 94010 Créteil,3 Laboratoire de Biologie Moléculaire, Hôpital Américain de Paris, Neuilly, France2

Received 27 November 2001/ Returned for modification 5 February 2002/ Accepted 29 March 2002

To improve the diagnosis of invasive aspergillosis (IA), we developed a LightCycler PCR assay targeted to Aspergillus fumigatus and A. flavus mitochondrial DNA. To avoid contamination, fully automated nucleic acid extraction with the MagNA Pure LC apparatus was used. The linearity of the results was achieved over a 6-log range of input A. fumigatus DNA, from 0.3 ng to 3 fg/10 µl of water. We retrospectively compared the LightCycler PCR and an enzyme-linked immunosorbent assay for the detection of galactomannan (GM) in serum from 14 patients with IA. The GM assay was more frequently positive (57 of 109; 52%) than the PCR assay (49 of 109; 45%). The LightCycler PCR assay, combined with automated DNA extraction, could be used in association with the GM assay to improve the reliability of IA diagnosis.

* Corresponding author. Mailing address: Laboratoire de Parasitologie-Mycologie, Hôpital Henri Mondor, 51 ave. du Maréchal DeLattre de Tassigny, 94010 Créteil Cedex, France. Phone: 33 1 49 81 36 41. Fax: 331 49 81 36 01. E-mail: bretagne{at}univ-paris12.fr.

Journal of Clinical Microbiology, June 2002, p. 2224-2227, Vol. 40, No. 6
0095-1137/02/$04.00+0     DOI: 10.1128/JCM.40.6.2224-2227.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Suarez, F., Lortholary, O., Buland, S., Rubio, M. T., Ghez, D., Mahe, V., Quesne, G., Poiree, S., Buzyn, A., Varet, B., Berche, P., Bougnoux, M. E. (2008). Detection of Circulating Aspergillus fumigatus DNA by Real-Time PCR Assay of Large Serum Volumes Improves Early Diagnosis of Invasive Aspergillosis in High-Risk Adult Patients under Hematologic Surveillance. J. Clin. Microbiol. 46: 3772-3777 [Abstract] [Full Text]  
  • Francesconi, A., Kasai, M., Harrington, S. M., Beveridge, M. G., Petraitiene, R., Petraitis, V., Schaufele, R. L., Walsh, T. J. (2008). Automated and Manual Methods of DNA Extraction for Aspergillus fumigatus and Rhizopus oryzae Analyzed by Quantitative Real-Time PCR. J. Clin. Microbiol. 46: 1978-1984 [Abstract] [Full Text]  
  • Barnes, R. A. (2008). Early diagnosis of fungal infection in immunocompromised patients. J Antimicrob Chemother 61: i3-i6 [Abstract] [Full Text]  
  • Kostamo, K., Richardson, M., Eerola, E., Rantakokko-Jalava, K., Meri, T., Malmberg, H., Toskala, E. (2007). Negative impact of Aspergillus galactomannan and DNA detection in the diagnosis of fungal rhinosinusitis. J Med Microbiol 56: 1322-1327 [Abstract] [Full Text]  
  • Baskova, L., Landlinger, C., Preuner, S., Lion, T. (2007). The Pan-AC assay: a single-reaction real-time PCR test for quantitative detection of a broad range of Aspergillus and Candida species. J Med Microbiol 56: 1167-1173 [Abstract] [Full Text]  
  • Schabereiter-Gurtner, C., Selitsch, B., Rotter, M. L., Hirschl, A. M., Willinger, B. (2007). Development of Novel Real-Time PCR Assays for Detection and Differentiation of Eleven Medically Important Aspergillus and Candida Species in Clinical Specimens. J. Clin. Microbiol. 45: 906-914 [Abstract] [Full Text]  
  • Mennink-Kersten, M. A. S. H., Ruegebrink, D., Wasei, N., Melchers, W. J. G., Verweij, P. E. (2006). In Vitro Release by Aspergillus fumigatus of Galactofuranose Antigens, 1,3-{beta}-D-Glucan, and DNA, Surrogate Markers Used for Diagnosis of Invasive Aspergillosis.. J. Clin. Microbiol. 44: 1711-1718 [Abstract] [Full Text]  
  • Espy, M. J., Uhl, J. R., Sloan, L. M., Buckwalter, S. P., Jones, M. F., Vetter, E. A., Yao, J. D. C., Wengenack, N. L., Rosenblatt, J. E., Cockerill, F. R. III, Smith, T. F. (2006). Real-Time PCR in Clinical Microbiology: Applications for Routine Laboratory Testing. Clin. Microbiol. Rev. 19: 165-256 [Abstract] [Full Text]  
  • Millon, L., Piarroux, R., Deconinck, E., Bulabois, C.-E., Grenouillet, F., Rohrlich, P., Costa, J.-M., Bretagne, S. (2005). Use of Real-Time PCR To Process the First Galactomannan-Positive Serum Sample in Diagnosing Invasive Aspergillosis. J. Clin. Microbiol. 43: 5097-5101 [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]  
  • 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]  
  • Elzi, L., Laifer, G., Bremerich, J., Vosbeck, J., Mayr, M. (2005). Invasive apergillosis with myocardial involvement after kidney transplantation. Nephrol Dial Transplant 20: 631-634 [Full Text]  
  • Singh, N., Paterson, D. L. (2005). Aspergillus Infections in Transplant Recipients. Clin. Microbiol. Rev. 18: 44-69 [Abstract] [Full Text]  
  • Musher, B., Fredricks, D., Leisenring, W., Balajee, S. A., Smith, C., Marr, K. A. (2004). Aspergillus Galactomannan Enzyme Immunoassay and Quantitative PCR for Diagnosis of Invasive Aspergillosis with Bronchoalveolar Lavage Fluid. J. Clin. Microbiol. 42: 5517-5522 [Abstract] [Full Text]  
  • Kawazu, M., Kanda, Y., Nannya, Y., Aoki, K., Kurokawa, M., Chiba, S., Motokura, T., Hirai, H., Ogawa, S. (2004). Prospective Comparison of the Diagnostic Potential of Real-Time PCR, Double-Sandwich Enzyme-Linked Immunosorbent Assay for Galactomannan, and a (1->3)-{beta}-D-Glucan Test in Weekly Screening for Invasive Aspergillosis in Patients with Hematological Disorders. J. Clin. Microbiol. 42: 2733-2741 [Abstract] [Full Text]  
  • Hsu, M.-C., Chen, K.-W., Lo, H.-J., Chen, Y.-C., Liao, M.-H., Lin, Y.-H., Li, S.-Y. (2003). Species identification of medically important fungi by use of real-time LightCycler PCR. J Med Microbiol 52: 1071-1076 [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]  


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