title: 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.
abstract: 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.
authors: Universidade do Estado do Amazonas, Manaus, Brazil. robertomsjr@hotmail.com.
authors: Roberto Moreira
authors: RM
authors: Da Silva
authors: Fundação de Medicina Tropical Dr. Heitor Vieira Dourado, Manaus, Brazil. netto.23@hotmail.com.
authors: João Ricardo
authors: JR
authors: Da Silva Neto
authors: Fundação de Medicina Tropical Dr. Heitor Vieira Dourado, Manaus, Brazil. carlasilvana@oi.com.br.
authors: Carla Silvana
authors: CS
authors: Santos
authors: Department of Tropical Medicine at the Bernhard Nocht Institute, German Armed Forces Hospital of Hamburg, Hamburg, Germany. frickmann@bni-hamburg.de.
authors: Hagen
authors: H
authors: Frickmann
authors: Institute of Medical Microbiology, Justus-Liebig-University Giessen, Giessen, Germany. svenpoppert@gmx.de.
authors: Sven
authors: S
authors: Poppert
authors: Fundação de Medicina Tropical Dr. Heitor Vieira Dourado, Manaus, Brazil. katia.cruz@fmt.am.gov.br.
authors: Kátia Santana
authors: KS
authors: Cruz
authors: Instituto de Criminalística, Manaus, Brazil. danielakoshikene@policiacivil.am.gov.br.
authors: Daniela
authors: D
authors: Koshikene
authors: Instituto Nacional de Pesquisas da Amazônia, Manaus, Brazil. joao.souza@inpa.gov.br.|Biotecnólogo/Tecnologista Pleno III, Instituto Nacional de Pesquisas da Amazônia, Coordenação de Sociedade, Ambiente e Saúde, Laboratório de Micologia, Av. André Araújo, 2936, Aleixo, Manaus, AM, CEP 69060-001, Brazil. joao.souza@inpa.gov.br.
authors: João Vicente Braga
authors: JV
authors: De Souza
mesh_terms: D000328:Adult; D001769:Blood / Q000382:microbiology*; D004271:DNA, Fungal / Q000235:genetics; D005260:Female; D005658:Fungi / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D017404:In Situ Hybridization, Fluorescence / Q000379:methods*; D008297:Male; D009181:Mycoses / Q000175:diagnosis / Q000382:microbiology*; D016133:Polymerase Chain Reaction
chemical_list: D004271:DNA, Fungal
references: J Appl Microbiol. 2011 Apr;110(4):881-92
references: 21205104
references: Braz J Med Biol Res. 2010 Aug;43(8):712-6
references: 20640387
references: Eur J Med Chem. 2010 Mar;45(3):1128-32
references: 20036039
references: Clin Microbiol Rev. 2011 Apr;24(2):247-80
references: 21482725
references: Curr Fungal Infect Rep. 2012 Mar;6(1):23-34
references: 22363832
references: J Clin Microbiol. 2011 Jun;49(6):2252-8
references: 21471340
references: J Clin Microbiol. 2013 Mar;51(3):863-8
references: 23269732
references: Eur J Clin Microbiol Infect Dis. 2010 May;29(5):571-6
references: 20221657
references: Nucl Med Biol. 2013 Jan;40(1):89-96
references: 23142409
references: J Clin Microbiol. 2007 Nov;45(11):3802-3
references: 17804657
references: J Clin Microbiol. 2008 Jan;46(1):50-5
references: 17977998
references: Infect Control Hosp Epidemiol. 2014 Aug;35(8):1077-9
references: 25026634
references: Mol Cell Probes. 1999 Dec;13(6):401-6
references: 10657143
references: J Bacteriol. 1990 Feb;172(2):762-70
references: 1688842
references: Rev Iberoam Micol. 2012 Jan-Mar;29(1):34-9
references: 21558013
references: Mycoses. 2013 Nov;56(6):681-6
references: 23560622
references: J Clin Microbiol. 2012 Apr;50(4):1446-8
references: 22238445
references: Mycoses. 2012 Sep;55(5):388-92
references: 22233292
references: J Clin Microbiol. 2006 Sep;44(9):3381-3
references: 16954279
references: Rev Soc Bras Med Trop. 2008 May-Jun;41(3):247-51
references: 18719803
references: Am J Clin Pathol. 2009 Mar;131(3):364-75
references: 19228642
references: J Clin Microbiol. 2000 Feb;38(2):830-8
references: 10655393
references: Med Mycol. 2015 Feb 1;53(2):160-4
references: 25537280
references: Mycoses. 2012 Nov;55(6):521-31
references: 22681159
references: Trends Microbiol. 2014 Mar;22(3):120-7
references: 24530175
references: J Clin Microbiol. 2006 Jan;44(1):119-23
references: 16390958
references: Mycoses. 2012 May;55(3):e114-23
references: 22126597
references: J Clin Microbiol. 2001 Oct;39(10):3578-82
references: 11574575
references: Antimicrob Agents Chemother. 1999 Feb;43(2):233-9
references: 9925511