A Comparison of Aspergillus and Mucorales PCR Testing of Different Bronchoalveolar Lavage Fluid Fractions from Patients with Suspected Invasive Pulmonary Fungal Disease

J Clin Microbiol. 2018 Jan 24;56(2):e01655-17. doi: 10.1128/JCM.01655-17. Print 2018 Feb.

Abstract

In patients with hematological malignancies, bronchoalveolar lavage fluid (BALF) specimens are commonly used for the diagnosis of mold infections. However, it is not clear whether the cell pellet (P) or the supernatant fraction (S) of the BALF specimen is optimal for molecular diagnostic testing. Thus, 99 BALF specimens were collected from 96 hematology patients with or without allogeneic hematopoietic stem cell transplant. The cell pellets and supernatants were processed alone and in combination (S/P) for testing by two fungus-specific real-time PCR assays compliant with international recommendations. The results achieved with S/P were revealed to be superior in comparison to those achieved with S and P alone, with the use of each single fraction showing a reduced sensitivity for the detection of Aspergillus DNA (82% and 43% for S and P, respectively). In 57% of the samples, testing of the combination of S and P generated a lower quantification cycle value than testing of S or P alone. Molds would have been missed in 5 and 16 out of 28 samples if only S or P, respectively, was analyzed. No sample was positive by testing of S or P only. Similar results were obtained for the detection of Mucorales DNA in BALF specimens (reduced sensitivity of 67% and 50% for S and P, respectively). Study patients were categorized according to the current European Organization for the Research and Treatment of Cancer/Mycoses Study Group classification for invasive fungal disease (IFD), revealing that 35 patients had proven/probable IFD (36%), 47 patients had possible IFD (49%), and 14 patients had undetermined IFD (15%).

Keywords: Aspergillus; Mucorales; PCR; bronchoalveolar lavage fluid; mold; pellet; supernatant.

Publication types

  • Comparative Study
  • Multicenter Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Aged
  • Aged, 80 and over
  • Aspergillus / genetics
  • Aspergillus / isolation & purification*
  • Bronchoalveolar Lavage Fluid / microbiology*
  • DNA, Fungal / genetics
  • Humans
  • Invasive Fungal Infections / diagnosis*
  • Invasive Pulmonary Aspergillosis / diagnosis
  • Middle Aged
  • Molecular Diagnostic Techniques / methods*
  • Molecular Diagnostic Techniques / standards
  • Mucorales / genetics
  • Mucorales / isolation & purification*
  • Real-Time Polymerase Chain Reaction* / standards
  • Sensitivity and Specificity
  • Young Adult

Substances

  • DNA, Fungal