title: tuf-PCR-temporal temperature gradient gel electrophoresis for molecular detection and identification of staphylococci: application to breast milk and neonate gut microbiota.
abstract: Coagulase negative staphylococci (CoNS) are a leading cause of infections in preterm infants, mostly involved in late-onset infection in low birth weight neonates. The epidemiology and pathophysiology of these infections remain unclear, notably because the causing agents are gathered in the artificial CoNS group. The aim of this work was to optimize the study of Staphylococcus species diversity in human breast milk and neonate stool, two sample types with bacterial communities dominated by CoNS, using PCR-temporal temperature gel electrophoresis based on the tuf gene. The optimized protocol identified 18 Staphylococcus species involved in neonate gut microbiota and infections and was applied to cultivation-independent study of breast milk and neonate stool. The efficiency, sensitivity, specificity and species discrimination of the proposed protocol appears suitable for patient follow-up in order to link microbiological data at the community level in milk and stool and interpret them from epidemiological and pathophysiological points of view.
authors.affiliation: Université Montpellier 1, UMR 5119 ECOSYM, Equipe Pathogènes et Environnements, U.F.R. des Sciences Pharmaceutiques et Biologiques, 15 Avenue Charles Flahault, BP 14491, 34093 Montpellier Cedex 5, France; Centre Hospitalier Régional Universitaire de Montpellier, Hôpital Lapeyronie, Département Urgences et Post-urgences Pédiatriques, 371 Avenue du Doyen Gaston Giraud, 34295 Montpellier Cedex 5, France.
authors.forename: Anne
authors.initials: A
authors.lastname: Filleron
authors.affiliation: Université Montpellier 1, UMR 5119 ECOSYM, Equipe Pathogènes et Environnements, U.F.R. des Sciences Pharmaceutiques et Biologiques, 15 Avenue Charles Flahault, BP 14491, 34093 Montpellier Cedex 5, France.
authors.forename: Margaux
authors.initials: M
authors.lastname: Simon
authors.affiliation: Université Montpellier 1, UMR 5119 ECOSYM, Equipe Pathogènes et Environnements, U.F.R. des Sciences Pharmaceutiques et Biologiques, 15 Avenue Charles Flahault, BP 14491, 34093 Montpellier Cedex 5, France.
authors.forename: Stefaniya
authors.initials: S
authors.lastname: Hantova
authors.affiliation: Centre Hospitalier Régional Universitaire de Montpellier, Hôpital Arnaud de Villeneuve, Service de Néonatalogie, 371 Avenue du Doyen Gaston Giraud, 34295 Montpellier Cedex 5, France.
authors.forename: Aurélien
authors.initials: A
authors.lastname: Jacquot
authors.affiliation: Centre Hospitalier Régional Universitaire de Montpellier, Hôpital Arnaud de Villeneuve, Service de Néonatalogie, 371 Avenue du Doyen Gaston Giraud, 34295 Montpellier Cedex 5, France.
authors.forename: Gilles
authors.initials: G
authors.lastname: Cambonie
authors.affiliation: Université Montpellier 1, UMR 5119 ECOSYM, Equipe Pathogènes et Environnements, U.F.R. des Sciences Pharmaceutiques et Biologiques, 15 Avenue Charles Flahault, BP 14491, 34093 Montpellier Cedex 5, France; Centre Hospitalier Régional Universitaire de Montpellier, Hôpital Arnaud de Villeneuve, Laboratoire de Bactériologie, 371 Avenue du Doyen Gaston Giraud, 34295 Montpellier Cedex 5, France.
authors.forename: Hélène
authors.initials: H
authors.lastname: Marchandin
authors.affiliation: Université Montpellier 1, UMR 5119 ECOSYM, Equipe Pathogènes et Environnements, U.F.R. des Sciences Pharmaceutiques et Biologiques, 15 Avenue Charles Flahault, BP 14491, 34093 Montpellier Cedex 5, France; Centre Hospitalier Régional Universitaire de Montpellier, Laboratoire d'Hygiène Hospitalière, 778 rue de la Croix Verte, 34000 Montpellier, France. Electronic address: ebilak@univ-montp1.fr.
authors.forename: Estelle
authors.initials: E
authors.lastname: Jumas-Bilak
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics*; D058645:Denaturing Gradient Gel Electrophoresis / Q000379:methods; D005243:Feces / Q000382:microbiology; D041981:Gastrointestinal Tract / Q000382:microbiology*; D006801:Humans; D007231:Infant, Newborn; D064307:Microbiota / Q000235:genetics; D008895:Milk, Human / Q000382:microbiology*; D016133:Polymerase Chain Reaction / Q000379:methods; D013203:Staphylococcal Infections / Q000175:diagnosis* / Q000382:microbiology; D013210:Staphylococcus / Q000302:isolation & purification*
chemical_list: D001426:Bacterial Proteins
keywords: Breast milk; Elongation factor Tu gene (tuf); Neonate; PCR-temporal temperature gel electrophoresis; Staphylococcus; Stool