title: Cultivation-independent approach for the direct detection of bacteria in human clinical specimens as a tool for analysing culture-negative samples: a prospective study.
abstract: Administration of empirical antibiotic therapy prior to microbiological diagnosis is thought to be associated the failure of subsequent bacterial growth in culture. The aim of this study was to detect bacterial pathogens via direct amplification and sequencing of the 16S rDNA gene in samples showing negative culture results as alternative diagnostic tools to troubleshoot difficult samples. Twenty-three (7.66 %) positive samples were detected, most of which were monomicrobial infections; 15 of the cases were identified as HAIs, 6 had catheter colonisation, and 2 had sample colonisation. The pathogens identified included Escherichia, Salmonella, Pseudomonas spp., Enterococcus spp. and coagulase-negative staphylococci (CoNS). The most frequent infections were bacteraemia and urinary tract infection, but meningitis, warm infection and soft tissue infection were also documented. These findings emphasise the efficacy and usefulness of molecular diagnosis, thus 16S rDNA gene analysis is strongly indicated by HAIs diagnostics.
authors.affiliation: Department of Microbiology, Instituto Politécnico Nacional (IPN), Escuela Nacional de Ciencias Biológicas (ENCB), Mexico City, D.F. Mexico.
authors.forename: Ma Guadalupe
authors.initials: MG
authors.lastname: Aguilera-Arreola
authors.affiliation: Department of Microbiology, Instituto Politécnico Nacional (IPN), Escuela Nacional de Ciencias Biológicas (ENCB), Mexico City, D.F. Mexico ; Microbial Genetic Resources Laboratory, Centro Nacional de Recursos Genéticos (CNRG)-INIFAP, Tepatitlán de Morelos, Jalisco Mexico.
authors.forename: Marcos Daniel
authors.initials: MD
authors.lastname: Martínez-Peña
authors.affiliation: Department of Microbiology, Instituto Politécnico Nacional (IPN), Escuela Nacional de Ciencias Biológicas (ENCB), Mexico City, D.F. Mexico ; Department of Zoology, Escuela Nacional de Ciencias Biológicas - Instituto Politécnico Nacional, Mexico City, D.F. Mexico.
authors.forename: Fabiola
authors.initials: F
authors.lastname: Hernández-Martínez
authors.affiliation: Special Test Laboratory, Centro Médico Nacional (CMN) 20 de Noviembre - Instituto de Seguridad y Servicios Sociales de los Trabajadores del Estado (ISSSTE), Mexico City, D.F. Mexico.
authors.forename: Sara R
authors.initials: SR
authors.lastname: Juárez Enriques
authors.affiliation: Department of Epidemiology, Centro Médico Nacional (CMN) 20 de Noviembre del Instituto de Seguridad y Servicios Sociales de los Trabajadores del Estado (ISSSTE), Mexico City, D.F. Mexico.
authors.forename: Beatriz
authors.initials: B
authors.lastname: Rico Verdín
authors.affiliation: Special Test Laboratory, Centro Médico Nacional (CMN) 20 de Noviembre - Instituto de Seguridad y Servicios Sociales de los Trabajadores del Estado (ISSSTE), Mexico City, D.F. Mexico.
authors.forename: Cristina
authors.initials: C
authors.lastname: Majalca-Martínez
authors.affiliation: Department of Microbiology, Instituto Politécnico Nacional (IPN), Escuela Nacional de Ciencias Biológicas (ENCB), Mexico City, D.F. Mexico.
authors.forename: Graciela
authors.initials: G
authors.lastname: Castro-Escarpulli
authors.affiliation: Department of Epidemiology, Centro Médico Nacional (CMN) 20 de Noviembre del Instituto de Seguridad y Servicios Sociales de los Trabajadores del Estado (ISSSTE), Mexico City, D.F. Mexico.
authors.forename: Enrique
authors.initials: E
authors.lastname: Albarrán-Fernández
authors.affiliation: Special Test Laboratory, Centro Médico Nacional (CMN) 20 de Noviembre - Instituto de Seguridad y Servicios Sociales de los Trabajadores del Estado (ISSSTE), Mexico City, D.F. Mexico.
authors.forename: S Cecilia
authors.initials: SC
authors.lastname: Serrano-López
keywords: 16S rDNA; Broad-range PCR; Culture-negative; Direct sequencing; Hospital-acquired infections