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

  • E. Fréalle
  • K. Decrucq
  • F. Botterel
  • B. Bouchindhomme
  • D. Camus
  • E. Dei-Cas
  • J. M. Costa
  • I. Yakoub-Agha
  • S. Bretagne
  • L. Delhaes


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.



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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Copyright information

© Springer-Verlag 2008

Authors and Affiliations

  • E. Fréalle
    • 1
  • K. Decrucq
    • 1
  • F. Botterel
    • 2
  • B. Bouchindhomme
    • 3
  • D. Camus
    • 1
  • E. Dei-Cas
    • 1
  • J. M. Costa
    • 2
    • 4
  • I. Yakoub-Agha
    • 5
  • S. Bretagne
    • 2
  • L. Delhaes
    • 1
  1. 1.EA 3609, Département de Parasitologie-Mycologie, Faculté de Médecine, Pôle de MicrobiologieCHRU de Lille, Université de Lille 2, Laboratoire d’Ecologie du Parasitisme, Institut Pasteur de LilleLilleFrance
  2. 2.Laboratoire de Parasitologie MycologieHôpital Henri Mondor APHP et Université Paris 12CréteilFrance
  3. 3.Département de PathologieCHRU de Lille, Université de Lille 2LilleFrance
  4. 4.Laboratoire Pasteur-CerbaCergy-PontoiseFrance
  5. 5.EA 2686, Service des Maladies du SangCHRU de Lille, Université de Lille 2LilleFrance

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