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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Publicado en:Biomédica: Revista del Instituto Nacional de Salud Vol. 43; pp. 255 - 267
Autores principales: Almeida-Silva, Fernando, Almeida-Paes, Rodrigo, Serra-Damasceno, Lisandra, Motta-Santos, Edwiges, Claudio Ferreira, Luiz, Pereira-Quintella, Leonardo, de Faria Ferreira, Marcela, de Medeiros-Muniz, Mauro, Zancopé-Oliveira, Rosely M.
Formato: Artículo
Publicado: Instituto Nacional de Salud of Colombia 2023 Supplement
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Acceso en línea:Ver este registro en EBSCOhost
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      dt: 2023 Supplement
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        atl: The conventional diagnosis challenge: Real-time PCR and nested PCR correlation with the scoring system for individuals at high-risk of Pneumocystis jirovecii pneumonia.
      aug:
        au:
          Almeida-Silva, Fernando
          Almeida-Paes, Rodrigo
          Serra-Damasceno, Lisandra
          Motta-Santos, Edwiges
          Claudio Ferreira, Luiz
          Pereira-Quintella, Leonardo
          de Faria Ferreira, Marcela
          de Medeiros-Muniz, Mauro
          Zancopé-Oliveira, Rosely M.
        affil:
          Laboratório de Micologia, Instituto Nacional de Infectologia Evandro Chagas, Fundação Oswaldo Cruz, Rio de Janeiro, Brasil
          Hospital São José de Doenças Infecciosas, Fortaleza, Ceará, Brasil
          Departamento de Saúde Comunitária, Faculdade de Medicina, Universidade Federal do Ceará, Fortaleza, Ceará, Brasil
          Laboratório de Medicina Intensiva, Instituto Nacional de Infectologia Evandro Chagas, Fundação Oswaldo Cruz, Rio de Janeiro, Brasil
          Serviço de Anatomia Patológica, Instituto Nacional de Infectologia Evandro Chagas, Fundação Oswaldo Cruz, Rio de Janeiro, Brasil
          Serviço Ambulatorial do Instituto Nacional de Infectologia Evandro Chagas, Fundação Oswaldo Cruz, Rio de Janeiro, Brasil
      su:
        Pneumocystis pneumonia
        Pneumocystis jiroveci
        Fluorescent antibody technique
        HIV-positive persons
        CD4 lymphocyte count
        AIDS patients
      sug:
        subj:
          Pneumocystis pneumonia
          Pneumocystis jiroveci
          Fluorescent antibody technique
          HIV-positive persons
          CD4 lymphocyte count
          AIDS patients
      keyword:
        direct
        fluorescent antibody technique
        neumonía por Pneumocystis
        pneumocystis
        Pneumonia
        reacción en cadena en tiempo real de la polimerasa
        real-time polymerase chain reaction
        técnica de inmunofluorescencia directa
        neumonía por Pneumocystis
        reacción en cadena en tiempo real de la polimerasa
        técnica de inmunofluorescencia directa
      ab:
        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.
        Introducción. Pneumocystis jirovecii es un hongo oportunista que afecta principalmente a personas con HIV (recuento de CD4 menor de 200 células/ml) y a otros pacientes inmunosuprimidos. Como P. jirovecii no crece en los medios micológicos de rutina, el diagnóstico de neumonía por P. jirovecii se basa en la evidencia presente en muestras respiratorias. Objetivos. Asociar los resultados de la inmunofluorescencia directa y los de dos métodos moleculares con un puntaje para predecir la neumonía causada por P. jirovecii en pacientes con sida. Materiales y métodos. Se realizó un estudio prospectivo de 40 pacientes. Se recolectó una muestra respiratoria antes del inicio de tratamiento y se sometió a una prueba de inmunofluorescencia directa con el kit Merifluor, una PCR anidada para la amplificación de la subunidad larga del ribosoma mitocondrial y una PCR en tiempo real usando el kit VIASURE. Resultados. Estas tres técnicas evidenciaron la presencia de P. jirovecii en 6, 12 y 15 muestras, respectivamente. Todas las muestras positivas por inmunofluorescencia directa fueron positivas en la PCR anidada y todas las muestras positivas en la PCR anidada amplificaron por PCR en tiempo real. Se encontró una asociación estadística entre los valores de la neumonía causada por P. jirovecii y los métodos moleculares. Dos pacientes con diagnóstico temprano respondieron satisfactoriamente al tratamiento. Conclusión. Se recomiendan los métodos moleculares, especialmente la PCR en tiempo real, para el diagnóstico temprano de neumonía causada por P. jirovecii en pacientes con sida.
      pubtype: Academic Journal
      doctype: Article
      src: R
    language: English
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