Use of Aspergillus fumigatus real-time PCR in bronchoalveolar lavage samples (BAL) for diagnosis of invasive aspergillosis, including azole-resistant cases, in high risk haematology patients: The need for a combined use with galactomannan

Malgorzata Mikulska, Elisa Furfaro, Elena De Carolis, Enrico Drago, Ilaria Pulzato, Maria Lucia Borghesi, Emanuela Zappulo, Anna Maria Raiola, Carmen Di Grazia, Valerio Del Bono, Giuseppe Cittadini, Emanuele Angelucci, Maurizio Sanguinetti, Claudio Viscoli

Research output: Contribution to journalArticle

Abstract

Diagnosis of invasive aspergillosis (IA) is challenging, particularly in high-risk patients with lung lesions other than typical according to 2008-EORTC/MSG criteria. Even if microbiology is positive, they still remain unclassified according to 2008-EORTC/MSG. Quantitative polymerase chain reaction (qPCR) provides new mycological documentation of IA. This retrospective study assessed Aspergillus fumigatus real time qPCR (MycoGENIE®R) in BAL to diagnose IA and identify azole-resistant strains. Clinical, radiological, and microbiological data from 114 hematology patients (69% HSCT recipients; 29% on mould active agents) from years 2012-2017 were collected; and 123 BAL samples were tested with qPCR (cutoff: Ct < 40) and galactomannan (GM, Platelia®R, cutoff: 0.5 ODI). Patients were classified as proven/probable, possible, and no-IA. “Atypical-IA” referred to patients with lesions other than typical according to 2008-EORTC/MSG and positive mycology. Proven IA was diagnosed in two cases (1.6%), probable in 28 (22.8%), possible in 27 (22%), atypical in 14 (11.4%). qPCR was positive in 39 samples (31.7%). Sensitivity and specificity of qPCR for proven/probable IA (vs no-IA; atypical-IA excluded) were 40% (95% confidence interval [CI]: 23-59) and 69% (95%CI: 55-81), respectively. Sensitivity of qPCR was higher when combined with GM (83%, 95%CI: 65-94) and in those receiving mould-active agents at BAL (61%, 95%CI: 32-86). One sample had TR34/L98H mutation. In conclusion, in high-risk hematology patients with various lung lesions, A. fumigatus qPCR in BAL contributes to diagnosing IA, particularly if combined with GM and in patients receiving mould-active agents might allow detecting azole-resistant mutations in culture negative samples.

Original languageEnglish
Pages (from-to)987-996
Number of pages10
JournalMedical Mycology
Volume57
Issue number8
DOIs
Publication statusPublished - Nov 1 2019

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Azoles
Aspergillus fumigatus
Aspergillosis
Bronchoalveolar Lavage
Hematology
Real-Time Polymerase Chain Reaction
Sodium Glutamate
Polymerase Chain Reaction
Confidence Intervals
Fungi
galactomannan
Mycology
Lung
Mutation
Microbiology
Documentation
Retrospective Studies
Sensitivity and Specificity

Keywords

  • Aspergillus fumigatus PCR
  • BAL
  • Galactomannan
  • HSCT
  • Invasive aspergillosis

ASJC Scopus subject areas

  • Infectious Diseases

Cite this

Use of Aspergillus fumigatus real-time PCR in bronchoalveolar lavage samples (BAL) for diagnosis of invasive aspergillosis, including azole-resistant cases, in high risk haematology patients : The need for a combined use with galactomannan. / Mikulska, Malgorzata; Furfaro, Elisa; De Carolis, Elena; Drago, Enrico; Pulzato, Ilaria; Borghesi, Maria Lucia; Zappulo, Emanuela; Raiola, Anna Maria; Grazia, Carmen Di; Bono, Valerio Del; Cittadini, Giuseppe; Angelucci, Emanuele; Sanguinetti, Maurizio; Viscoli, Claudio.

In: Medical Mycology, Vol. 57, No. 8, 01.11.2019, p. 987-996.

Research output: Contribution to journalArticle

Mikulska, Malgorzata ; Furfaro, Elisa ; De Carolis, Elena ; Drago, Enrico ; Pulzato, Ilaria ; Borghesi, Maria Lucia ; Zappulo, Emanuela ; Raiola, Anna Maria ; Grazia, Carmen Di ; Bono, Valerio Del ; Cittadini, Giuseppe ; Angelucci, Emanuele ; Sanguinetti, Maurizio ; Viscoli, Claudio. / Use of Aspergillus fumigatus real-time PCR in bronchoalveolar lavage samples (BAL) for diagnosis of invasive aspergillosis, including azole-resistant cases, in high risk haematology patients : The need for a combined use with galactomannan. In: Medical Mycology. 2019 ; Vol. 57, No. 8. pp. 987-996.
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abstract = "Diagnosis of invasive aspergillosis (IA) is challenging, particularly in high-risk patients with lung lesions other than typical according to 2008-EORTC/MSG criteria. Even if microbiology is positive, they still remain unclassified according to 2008-EORTC/MSG. Quantitative polymerase chain reaction (qPCR) provides new mycological documentation of IA. This retrospective study assessed Aspergillus fumigatus real time qPCR (MycoGENIE{\circledR}R) in BAL to diagnose IA and identify azole-resistant strains. Clinical, radiological, and microbiological data from 114 hematology patients (69{\%} HSCT recipients; 29{\%} on mould active agents) from years 2012-2017 were collected; and 123 BAL samples were tested with qPCR (cutoff: Ct < 40) and galactomannan (GM, Platelia{\circledR}R, cutoff: 0.5 ODI). Patients were classified as proven/probable, possible, and no-IA. “Atypical-IA” referred to patients with lesions other than typical according to 2008-EORTC/MSG and positive mycology. Proven IA was diagnosed in two cases (1.6{\%}), probable in 28 (22.8{\%}), possible in 27 (22{\%}), atypical in 14 (11.4{\%}). qPCR was positive in 39 samples (31.7{\%}). Sensitivity and specificity of qPCR for proven/probable IA (vs no-IA; atypical-IA excluded) were 40{\%} (95{\%} confidence interval [CI]: 23-59) and 69{\%} (95{\%}CI: 55-81), respectively. Sensitivity of qPCR was higher when combined with GM (83{\%}, 95{\%}CI: 65-94) and in those receiving mould-active agents at BAL (61{\%}, 95{\%}CI: 32-86). One sample had TR34/L98H mutation. In conclusion, in high-risk hematology patients with various lung lesions, A. fumigatus qPCR in BAL contributes to diagnosing IA, particularly if combined with GM and in patients receiving mould-active agents might allow detecting azole-resistant mutations in culture negative samples.",
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AU - Mikulska, Malgorzata

AU - Furfaro, Elisa

AU - De Carolis, Elena

AU - Drago, Enrico

AU - Pulzato, Ilaria

AU - Borghesi, Maria Lucia

AU - Zappulo, Emanuela

AU - Raiola, Anna Maria

AU - Grazia, Carmen Di

AU - Bono, Valerio Del

AU - Cittadini, Giuseppe

AU - Angelucci, Emanuele

AU - Sanguinetti, Maurizio

AU - Viscoli, Claudio

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