Performance of a Real Time PCR for Pneumocystis jirovecii Identification in Induced Sputum of AIDS Patients: Differentiation between Pneumonia and Colonization.

Autor: Chagas OJ; Laboratório de Investigação Médica em Micologia LIM53, Instituto de Medicina Tropical e Hospital das Clínicas, Faculdade de Medicina da Universidade de São Paulo, São Paulo 05403-000, Brazil., Nagatomo PP; Laboratório de Investigação Médica em Micologia LIM53, Instituto de Medicina Tropical e Hospital das Clínicas, Faculdade de Medicina da Universidade de São Paulo, São Paulo 05403-000, Brazil., Pereira-Chioccola VL; Laboratório de Biologia Molecular de Parasitas e Fungos do Centro de Parasitologia e Micologia, Instituto Adolfo Lutz, São Paulo 01246-000, Brazil., Gava R; Laboratório de Biologia Molecular de Parasitas e Fungos do Centro de Parasitologia e Micologia, Instituto Adolfo Lutz, São Paulo 01246-000, Brazil., Buccheri R; Instituto de Infectologia Emílio Ribas, São Paulo 01246-900, Brazil., Del Negro GMB; Laboratório de Investigação Médica em Micologia LIM53, Instituto de Medicina Tropical e Hospital das Clínicas, Faculdade de Medicina da Universidade de São Paulo, São Paulo 05403-000, Brazil., Benard G; Laboratório de Investigação Médica em Micologia LIM53, Instituto de Medicina Tropical e Hospital das Clínicas, Faculdade de Medicina da Universidade de São Paulo, São Paulo 05403-000, Brazil.
Jazyk: angličtina
Zdroj: Journal of fungi (Basel, Switzerland) [J Fungi (Basel)] 2022 Feb 24; Vol. 8 (3). Date of Electronic Publication: 2022 Feb 24.
DOI: 10.3390/jof8030222
Abstrakt: Pneumocystis jirovecii pneumonia (PcP) remains an important cause of morbimortality worldwide and a diagnostic challenge. Conventional methods have low accuracy, hardly discriminating colonization from infection, while some new high-cost or broncho-alveolar lavage-based methods have limited usefulness in developing countries. Quantitative PCR (qPCR) tests may overcome these limitations due to their high accuracy, possibility of automation, and decreasing cost. We evaluated an in-house qPCR targeting the fungus mtSSU gene using induced sputum. Sensitivity of the assay (ten target gene copies/assay) was determined using recombinant plasmids. We prospectively studied 86 AIDS patients with subacute respiratory symptoms in whom PcP was suspected. qPCR results were determined as quantification cycles (Cq) and compared with a qualitative PCR performed in the same IS, serum 1,3-β-D-Glucan assay, and a clinical/laboratory/radiology index for PcP. The qPCR clustered the patients in three groups: 32 with Cq ≤ 31 (qPCR+), 45 with Cq ≥ 33 (qPCR-), and nine with Cq between 31-33 (intermediary), which, combined with the other three analyses, enabled us to classify the groups as having PcP, not P. jirovecii -infected, and P. jirovecii -colonized, respectively. This molecular assay may contribute to improve PcP management, avoiding unnecessary treatments, and our knowledge of the natural history of this infection.
Databáze: MEDLINE
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