title: Use of calcofluor-blue brightener for the diagnosis of Pneumocystis jirovecii pneumonia in bronchial-alveolar lavage fluids: A single-center prospective study.
abstract: The biological diagnosis of Pneumocystis jirovecii pneumonia (PjP) is based on the investigation of respiratory fluids by conventional staining methods and/or molecular biology. Diagnostic performance of an in-house technique based on calcofluor-blue brightener for the direct detection of P. jirovecii cysts was prospectively assessed in bronchial-alveolar lavage fluids (BALF) from patients with a suspected PjP infection over a three-year period in a single center: the diagnostic yield was compared to that of a commercial kit based on monoclonal immunofluorescence assay (IFA) on replicate smears. May-Grünwald Giemsa (MGG) staining and quantitative Polymerase Chain Reaction (qPCR) were also performed. The gold standard for each patient was the definitive diagnosis of PjP infection by an independent committee based on clinical, radiological, and biological data. Overall, 481 BALF were assessed: 42 were found to be positive for the detection of P. jirovecii by at least one laboratory technique, but only 35 were actually judged to be in agreement with the definitive diagnosis of PjP infection. The sensitivity of the calcofluor-blue brightener technique was 74.3% vs. 60.0%, 34.6%, and 82.9% for IFA, MGG, and qPCR, respectively; and its specificity was 99.6% vs. 99.3%, 100.0%, and 99.4% for IFA, MGG, and qPCR. No technique was shown to be statistically superior to calcofluor-blue brightener. Further validation of the test through multicenter studies is now required, but in light of its low cost and easy preparation, the use of calcofluor-blue brightener in BALF appears to be a valuable alternative method for the routine first-line diagnosis of PjP infection.
authors: CHU de Tours, Service de Parasitologie - Mycologie - Médecine tropicale, Tours - France.|Université François-Rabelais, CEPR - INSERM U1100 / Équipe 3, Faculté de Médecine, Tours - France.
authors: Guillaume
authors: G
authors: Desoubeaux
authors: CHU de Tours, Service de Parasitologie - Mycologie - Médecine tropicale, Tours - France.
authors: Claire
authors: C
authors: Franck-Martel
authors: CHU de Tours, CIC - INSERM 1415, Faculté de Médecine, Tours - France.|Université François-Rabelais, PRES Centre-Val de Loire, Tours - France.
authors: Agnès
authors: A
authors: Caille
authors: CHU de Tours, Service de Parasitologie - Mycologie - Médecine tropicale, Tours - France.
authors: Nicolas
authors: N
authors: Drillaud
authors: CHU de Tours, Service de Parasitologie - Mycologie - Médecine tropicale, Tours - France.|Université François-Rabelais, CEPR - INSERM U1100 / Équipe 3, Faculté de Médecine, Tours - France.
authors: Marie-Alix
authors: MA
authors: Lestrade Carluer de Kyvon
authors: CHU de Tours, Service de Parasitologie - Mycologie - Médecine tropicale, Tours - France.
authors: Éric
authors: É
authors: Bailly
authors: CHU de Tours, Service de Parasitologie - Mycologie - Médecine tropicale, Tours - France.|Université François-Rabelais, CEPR - INSERM U1100 / Équipe 3, Faculté de Médecine, Tours - France.
authors: Jacques
authors: J
authors: Chandenier
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D001992:Bronchoalveolar Lavage Fluid / Q000382:microbiology*; D005260:Female; D005456:Fluorescent Dyes / Q000378:metabolism*; D006801:Humans; D008297:Male; D008875:Middle Aged; D045363:Pneumocystis carinii / Q000302:isolation & purification*; D011020:Pneumonia, Pneumocystis / Q000175:diagnosis* / Q000382:microbiology*; D011446:Prospective Studies; D012680:Sensitivity and Specificity; D013194:Staining and Labeling / Q000379:methods*; D055815:Young Adult
chemical_list: D005456:Fluorescent Dyes
keywords: Pneumocystis jirovecii; Uvitex; bronchial-alveolar lavage; calcofluor-blue; immunofluorescence