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Diagnosis of invasive aspergillosis using bronchoalveolar lavage in haematology patients: influence of bronchoalveolar lavage human DNA content on real-time PCR performance

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Abstract

In order to improve invasive pulmonary aspergillosis (IPA) diagnosis, a real-time polymerase chain reaction (PCR) assay detecting Aspergillus spp. was developed. Its detection limit reached 2–20 conidia. The retrospective evaluation on 64 bronchoalveolar lavage (BAL) fluids from 57 patients at risk for IPA, including 20 probable and five proven IPA patients, revealed a 88% or 38% sensitivity in direct examination (DE)/culture-positive or culture-negative BAL, respectively, whereas galactomannan (GM) sensitivity reached 88% or 58%, respectively. Influence on the Aspergillus-PCR yield of BAL fluid volume, cellular count and DNA content (evaluated by human β-globin quantification) was assessed. Significantly higher β-globin levels were detected in Aspergillus PCR-positive (median 5,112 pg/μl) than negative (median 1,332 pg/μl) BAL fluids, suggesting that the β-globin level could reflect BAL yields and DNA extraction. Using β-globin for the interpretation of fungal PCR could improve the negative predictive value of this test.

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Acknowledgements

This work was supported by grants from the Lille 2 University Hospital of Lille and the Faculty of Medicine (Lille 2 University), and was developed in the framework of the EA 3609 scientific project (French Research Office). It was presented at the 3rd Trends in Medical Mycology (TIMM 2007), Turin, Italy, October 2007. We thank the members of the Department of Parasitology-Mycology of Lille 2 University Hospital: Fabienne Soula for her valuable help in the clinical data collection; Michèle Wauquier and Filoména Naji for their technical assistance; Boualem Sendid, Nadine François, Stéphanie Delbart, Pascale Daelman and Laurence Dumortier for their involvement in the routine mycological tests. We are grateful to Anthony Pinon (research assistant, Lille Pasteur Institute, France) for the statistical analysis.

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Authors and Affiliations

  1. EA 3609, Département de Parasitologie-Mycologie, Faculté de Médecine, Pôle de Microbiologie, CHRU de Lille, Université de Lille 2, Laboratoire d’Ecologie du Parasitisme, Institut Pasteur de Lille, Lille, France

    E. Fréalle, K. Decrucq, D. Camus, E. Dei-Cas & L. Delhaes

  2. Laboratoire de Parasitologie Mycologie, Hôpital Henri Mondor APHP et Université Paris 12, Créteil, France

    F. Botterel, J. M. Costa & S. Bretagne

  3. Département de Pathologie, CHRU de Lille, Université de Lille 2, Lille, France

    B. Bouchindhomme

  4. Laboratoire Pasteur-Cerba, Cergy-Pontoise, France

    J. M. Costa

  5. EA 2686, Service des Maladies du Sang, CHRU de Lille, Université de Lille 2, Lille, France

    I. Yakoub-Agha

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  1. E. Fréalle
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  2. K. Decrucq
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  9. S. Bretagne
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Correspondence to E. Fréalle.

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Fréalle, E., Decrucq, K., Botterel, F. et al. Diagnosis of invasive aspergillosis using bronchoalveolar lavage in haematology patients: influence of bronchoalveolar lavage human DNA content on real-time PCR performance. Eur J Clin Microbiol Infect Dis 28, 223–232 (2009). https://doi.org/10.1007/s10096-008-0616-1

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