The conventional diagnosis challenge: Real-time PCR and nested PCR correlation with the scoring system for individuals at high-risk of Pneumocystis jirovecii pneumonia

Introduction. Pneumocystis jirovecii is an opportunistic fungus that affects mainly people living with HIV (CD4 cell count lower than 200 cells/ml) and other immunosuppressed patients. Since P. jirovecii does not grow on routine mycological media, diagnosis of P. jirovecii pneumonia relies on indire...

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Bibliographic Details
Published in:Biomédica
Main Authors: Fernando Almeida-Siva, Rodrigo Almeida-Paes, Lisandra Serra-Damasceno, Edwiges Motta-Santos, Luiz Claudio Ferreira, Leonardo Pereira-Quintella, Marcela de Faria Ferreira, Mauro de Medeiros-Muniz, Rosely M. Zancopé-Oliveira
Format: Article in Journal/Newspaper
Language:English
Spanish
Published: Instituto Nacional de Salud 2023
Subjects:
pneumonia
pneumocystis
fluorescent antibody technique
direct
real-time polymerase chain reaction
Medicine
R
Arctic medicine. Tropical medicine
RC955-962
Arctic
Online Access:https://doi.org/10.7705/biomedica.7020
https://doaj.org/article/35517f35513d459e9af03adb6731d045
Description
Summary:Introduction. Pneumocystis jirovecii is an opportunistic fungus that affects mainly people living with HIV (CD4 cell count lower than 200 cells/ml) and other immunosuppressed patients. Since P. jirovecii does not grow on routine mycological media, diagnosis of P. jirovecii pneumonia relies on indirect evidence of its presence in respiratory samples. Objectives. To associate the results of direct immunofluorescence and two molecular methods with a score to predict P. jirovecii pneumonia in patients with AIDS. Materials and methods. A prospective study was conducted with 40 patients. A respiratory sample collected before treatment was subjected to direct immunofluorescence using the Merifluor kit, to nested PCR targeting the mitochondrial large subunit ribosomal RNA, and to the VIASURE real-time PCR kit. Results. These three techniques revealed P. jirovecii in 6, 12, and 15 samples, respectively. All positive samples by direct immunofluorescence were positive by nested PCR, and all positive samples by nested PCR amplified by real-time PCR. There was a statistically significant association between the P. jirovecii pneumonia score and the molecular methods. Two patients were early diagnosed and responded well to treatment. Conclusion. Molecular methods, especially real-time PCR, are recommended for early diagnosis of P. jirovecii pneumonia in AIDS patients.

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