title: Aspergillus-Specific Lateral-Flow Device and Real-Time PCR Testing of Bronchoalveolar Lavage Fluid: a Combination Biomarker Approach for Clinical Diagnosis of Invasive Pulmonary Aspergillosis.
abstract: Clinical experience with the impact of serum biomarkers for invasive fungal disease (IFD) varies markedly in hemato-oncology. Invasive pulmonary aspergillosis (IPA) is the most common manifestation, so we evaluated biomarkers in bronchoalveolar lavage (BAL) fluid. An Aspergillus-specific lateral-flow device (LFD), quantitative real-time PCR (qPCR), and the galactomannan (GM) test were used with 32 BAL fluid samples from 32 patients at risk of IPA. Eight patients had proven IPA, 3 had probable IPA, 6 had possible IPA, and 15 patients had no IPA by European Organization for Research and Treatment of Cancer Invasive Fungal Infections Cooperative Group/Mycoses Study Group of the National Institute of Allergy and Infectious Diseases (EORTC/MSG) criteria. The diagnostic accuracies of the tests were evaluated, and pairwise agreement between biomarkers was calculated. The diagnostic performance of the EORTC/MSG criteria was evaluated against the test(s) identified to be the most useful for IPA diagnosis. Using the EORTC/MSG criteria, the sensitivities of qPCR and LFD were 100% and the sensitivity of the GM test was 87.5% (GM test index cutoff, >0.8), with the tests having specificities of between 66.7 and 86.7%. The agreement between the results of qPCR and LFD was almost perfect (Cohen's kappa coefficient = 0.93, 95% confidence interval, 0.81 to 1.00). LFD and qPCR combined had a sensitivity of 100% and a specificity of 85.7%. Calcofluor staining and culture of all BAL fluid samples were negative for fungal infection. The median time from the start of mold-active antifungal therapy to the time of collection of BAL fluid was 6 days. Reversing roles and using dual testing by LFD and qPCR to classify cases, the EORTC/MSG criteria had a sensitivity of 83.3%. All three tests are useful for the diagnosis of IPA in BAL fluid samples. Despite the significant delays between the start of antifungal therapy and bronchoscopy, unlike microscopy and culture, the biomarkers remained informative. In particular, the combination of LFD and qPCR allows the sensitive and specific detection of IPA.
authors.affiliation: Blizard Institute of Cell and Molecular Science, Queen Mary University of London, London, United Kingdom gemma.johnson@bartshealth.nhs.uk s.g.agrawal@qmul.ac.uk.
authors.forename: Gemma L
authors.initials: GL
authors.lastname: Johnson
authors.affiliation: Centre for Experimental Cancer Medicine, Barts Cancer Institute, Queen Mary University of London, London, United Kingdom.
authors.forename: Shah-Jalal
authors.initials: SJ
authors.lastname: Sarker
authors.affiliation: Blizard Institute of Cell and Molecular Science, Queen Mary University of London, London, United Kingdom.
authors.forename: Francesco
authors.initials: F
authors.lastname: Nannini
authors.affiliation: Blizard Institute of Cell and Molecular Science, Queen Mary University of London, London, United Kingdom.
authors.forename: Arianna
authors.initials: A
authors.lastname: Ferrini
authors.affiliation: Public Health Laboratory London, Public Health England, London, United Kingdom.
authors.forename: Emma
authors.initials: E
authors.lastname: Taylor
authors.affiliation: Division of Hygiene and Medical Microbiology, Innsbruck Medical University, Innsbruck, Austria.
authors.forename: Cornelia
authors.initials: C
authors.lastname: Lass-Flörl
authors.affiliation: Division of Hygiene and Medical Microbiology, Innsbruck Medical University, Innsbruck, Austria.
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Mutschlechner
authors.affiliation: Postgraduate Medical Institute, Faculty of Medical Science, Anglia Ruskin University, Chelmsford, United Kingdom.
authors.forename: Stephen A
authors.initials: SA
authors.lastname: Bustin
authors.affiliation: Blizard Institute of Cell and Molecular Science, Queen Mary University of London, London, United Kingdom Department of Haemato-Oncology, St. Bartholomew's Hospital, London, United Kingdom gemma.johnson@bartshealth.nhs.uk s.g.agrawal@qmul.ac.uk.
authors.forename: Samir G
authors.initials: SG
authors.lastname: Agrawal
mesh_terms: D000328:Adult; D000946:Antigens, Fungal / Q000032:analysis / Q000276:immunology; D015415:Biomarkers / Q000032:analysis*; D001992:Bronchoalveolar Lavage Fluid / Q000382:microbiology*; D002846:Chromatography, Affinity / Q000379:methods*; D004271:DNA, Fungal / Q000032:analysis / Q000235:genetics; D006801:Humans; D055744:Invasive Pulmonary Aspergillosis / Q000175:diagnosis*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D012189:Retrospective Studies; D012680:Sensitivity and Specificity
chemical_list: D000946:Antigens, Fungal; D015415:Biomarkers; D004271:DNA, Fungal