Rapid and specific detection of section Fumigati and Aspergillus fumigatus in human samples using a new multiplex real-time PCR

Fecha de publicación: 01/10/2014

Autores de IIS La Fe

Javier Pemán García

Autor

Participantes ajenos a IIS La Fe

Fernandez-Molina JV
Abad-Diaz-de-Cerio A
Sueiro-Olivares M
Pellon A
Ramirez-Garcia A
Garaizar J
Hernando FL
Rementeria A

Grupos

Infección Grave

Investigador Principal

María Ángeles Tormo Mas

Abstract

Invasive aspergillosis is an opportunistic infection caused primarily by Aspergillus fumigatus. However, other common fungal pathogens belonging to section Fumigati are often misidentified as A. fumigatus. Thus, we have developed a multiplex real-time PCR (qPCR) assay with primers and specific TaqMan probes based on internal transcribed spacer regions or benA gene to discriminate, in less than 3 h, species of section Fumigati and, specifically, A. fumigatus. The multiplex qPCR showed a limit of detection of 20 and 50 fg of DNA for section Fumigati and A. fumigatus, respectively. Moreover, it enabled detection of a single germinated conidia. The inclusion of some PCR facilitators together with the dilution of samples makes it possible to completely avoid PCR inhibitions in all bronchoalveolar lavage (BAL) samples assayed. This technique may be a useful complementary tool in the diagnosis of invasive pulmonary aspergillosis caused by A. fumigatus using BAL fluid. (C) 2014 Elsevier Inc. All rights reserved.

Datos de la publicación

ISSN/ISSNe:: 0732-8893, 1879-0070
Tipo:: Article
Páginas:: 111-118
DOI:: 10.1016/j.diagmicrobio.2014.06.003
Factor de Impacto:: 1,203 SCImago ℠
Cuartil:: Q1 SCImago ℠

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Filiaciones

Fernandez-Molina JV:: Department of Immunology, Microbiology and Parasitology, Faculty of Science and Technology, University of the Basque Country (UPV/EHU), Spain
Abad-Diaz-de-Cerio A:: Department of Immunology, Microbiology and Parasitology, Faculty of Science and Technology, University of the Basque Country (UPV/EHU), Spain
Sueiro-Olivares M:: Department of Immunology, Microbiology and Parasitology, Faculty of Science and Technology, University of the Basque Country (UPV/EHU), Spain
Pellon A:: Department of Immunology, Microbiology and Parasitology, Faculty of Science and Technology, University of the Basque Country (UPV/EHU), Spain
Ramirez-Garcia A:: Department of Immunology, Microbiology and Parasitology, Faculty of Science and Technology, University of the Basque Country (UPV/EHU), Spain
Garaizar J:: Department of Immunology, Microbiology and Parasitology, Faculty of Pharmacy, University of the Basque Country (UPV/EHU), Spain
Pemán J:: Department of Microbiology, Hospital Universitario y Politécnico La Fe, Valencia, Spain
Hernando FL:: Department of Immunology, Microbiology and Parasitology, Faculty of Science and Technology, University of the Basque Country (UPV/EHU), Spain
Rementeria A:: Department of Immunology, Microbiology and Parasitology, Faculty of Science and Technology, University of the Basque Country (UPV/EHU), Spain.

Keywords

Aspergillus fumigatus; Section Fumigati; Multiplex real-time PCR; ITS region; benA gene; BAL samples

Proyectos asociados

FUNGEMIA: EPIDEMIOLOGIA, MECANISMOS DE RESISTENCIA ANTIFUNGICA Y TIPADO MOLECULAR

090/2006- EVES - PEMAN . 2006

NUEVAS ESTRATEGIAS PARA EL TRATAMIENTO DE LAS INFECCIONES ASOCIADAS A BIOCAPAS FUNGICAS.

Investigador Principal: EMILIA CANTON LACASA

PI12/02786 . INSTITUTO DE SALUD CARLOS III . 2013

ESTUDIO EPIDEMIOLOGICO DE DIAGNOSTICO PRECOZ DE LA CANDIDIASIS INVASORA EN PACIENTES NEONATOS PRETERMINO DE MUY BAJO PESO. ESTUDIO CANDI-NEO

Investigador Principal: MÁXIMO VENTO TORRES

AST-MYC-2010-01

ESTUDIO OBSERVACIONAL, PROSPECTIVO Y MULTICÉNTRICO EN CUIDADOS INTENSIVOS PARA EVALUAR LA FIABILIDAD DIAGNÓSTICOA DE LA PCR (REACCIÓN EN CADENA DE LA POLIMERASA) EN PACIENTES EN TRATAMIENTO ANTIFÚNGICO EMPÍRICO POR SOSPECHA DE CANDIDIASIS INVASIVA. (ESTUD IO MICAFEM).

Investigador Principal: PAULA RAMÍREZ GALLEYMORE

AST-INF-2012-01

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