Fluorescent in situ hybridization of pre-incubated blood culture material for the rapid diagnosis of histoplasmosis

• Autor(a) principal: Silva, Roberto Moreira da
• Data de Publicação: 2015
• Outros Autores: Silva Neto, João Ricardo da, Santos, Carla Silvana, Cruz, Kátia Santana, Frickmann, Hagen, Poppert, Sven, Koshikene, Daniela, Souza, João Vicente Braga de
• Tipo de documento: Artigo
• Idioma: eng
• Título da fonte: Repositório Institucional do INPA
• DOI: 10.1093/mmy/myu080
• Texto Completo: https://repositorio.inpa.gov.br/handle/1/15881
• topic: Fluorescent Dyes; Ribosome Rna; Fungal Rna; Oligonucleotide Probe; Ribosome Rna; Ajellomyces Capsulatus; Bacteria; Blood Culture; Clinical In Situ Hybridization, Fluorescence; Fungal Detection; Fungal Strain; Fungus Identification; Histoplasmosis; Human; Oligonucleotide Probe; Polymerase Chain Reaction; Yeast; Blood; In Situ Hybridization, Fluorescence; Genetics; Histoplasma; Histoplasmosis; Isolation And Purification; Microbiology; Molecular Diagnosis; Procedures; Sensitivity And Specificity; Time; Candida; Histoplasma Capsulatum Var. Capsulatum; Blood; Histoplasma; Histoplasmosis; Humans; In Situ Hybridization, Fluorescence; Molecular Diagnostic Techniques; Oligonucleotide Probes; Fungal Rna; Rna, Ribosomal; Sensitivity And Specificity; Time Factors

Fluorescence in situ hybridization (FISH) has been shown to be useful for the detection of Candida and Cryptococcus species in blood culture materials. FISH procedures for the detection of Histoplasma capsulatum var. capsulatum have not been reported so far. This study describes the development and evaluation of fluorescently labeled rRNA-targeting FISH probes to detect and identify H. capsulatum in blood cultures. All three analyzed H. capsulatum reference strains and clinical isolates showed positive signals with the newly designed specific oligonucleotide probes for H. capsulatum, whereas negative reactions were observed for all three nontarget yeast species and the two nontarget bacteria. The assay was also successfully applied for detections of H. capsulatum cells in pre-incubated blood culture samples of patients with clinical suspicion of histoplasmosis (n = 33). The described FISH-based assay was shown to be easy to apply, sensitive, and specific (compared to polymerase chain reaction) for the detection and identification of H. capsulatum in this proof-of-principle analysis. Larger multicentric assessments are recommended for a thorough diagnostic evaluation of the procedure. © The Author 2014. Published by Oxford University Press on behalf of The International Society for Human and Animal Mycology.