Evaluation of fluorescence in situ hybridisation (FISH) for the detection of fungi directly from blood cultures and cerebrospinal fluid from patients with suspected invasive mycoses

• Autor(a) principal: Silva, Roberto Moreira da
• Data de Publicação: 2015
• Outros Autores: Silva Neto, João Ricardo da, Santos, Carla Silvana, Frickmann, Hagen, Poppert, Sven, Cruz, Kátia Santana, Koshikene, Daniela, Souza, João Vicente Braga de
• Tipo de documento: Artigo
• Idioma: eng
• Título da fonte: Repositório Institucional do INPA
• Texto Completo: https://repositorio.inpa.gov.br/handle/1/15908
• topic: Adolescent; Adult; Blood Culture; Bloodstream Infection; Cerebrospinal Fluid; Child; Controlled Study; Cryptococcosis; Diagnostic Test Accuracy Study; Female; In Situ Hybridization, Fluorescence; Fungal Detection; Hospital Laboratory; Human; Major Clinical Study; Male; Microbial Growth; Polymorphism, Restriction Fragment Length; Sensitivity And Specificity; Systemic Mycosis; Blood; Classification; Evaluation Study; In Situ Hybridization, Fluorescence; Fungus; Genetics; Isolation And Purification; Microbiology; Mycoses; Polymerase Chain Reaction; Procedures; Dna, Fungal; Adult; Blood; Dna, Fungal; Female; Fungi; Humans; In Situ Hybridization, Fluorescence; Male; Mycoses; Polymerase Chain Reaction

The aim of this study was to evaluate the diagnostic performance of in-house FISH (fluorescence in situ hybridisation) procedures for the direct identification of invasive fungal infections in blood cultures and cerebrospinal fluid (CSF) samples and to compare these FISH results with those obtained using traditional microbiological techniques and PCR targeting of the ITS1 region of the rRNA gene. In total, 112 CSF samples and 30 positive blood cultures were investigated by microscopic examination, culture, PCR-RFLP and FISH. The sensitivity of FISH for fungal infections in CSF proved to be slightly better than that of conventional microscopy (India ink) under the experimental conditions, detecting 48 (instead of 46) infections in 112 samples. The discriminatory powers of traditional microbiology, PCR-RFLP and FISH for fungal bloodstream infections were equivalent, with the detection of 14 fungal infections in 30 samples. However, the mean times to diagnosis after the detection of microbial growth by automated blood culture systems were 5 hours, 20 hours and 6 days for FISH, PCR-RFLP and traditional microbiology, respectively. The results demonstrate that FISH is a valuable tool for the identification of invasive mycoses that can be implemented in the diagnostic routine of hospital laboratories. © 2015 Da Silva et al.