title: More Easily Cultivated Than Identified: Classical Isolation With Molecular Identification of Vaginal Bacteria.
abstract: Women with bacterial vaginosis (BV) have complex communities of anaerobic bacteria. There are no cultivated isolates of several bacteria identified using molecular methods and associated with BV. It is unclear whether this is due to the inability to adequately propagate these bacteria or to correctly identify them in culture.
  
  
  
  Vaginal fluid from 15 women was plated on 6 different media using classical cultivation approaches. Individual isolates were identified by 16S ribosomal RNA (rRNA) gene sequencing and compared with validly described species. Bacterial community profiles in vaginal samples were determined using broad-range 16S rRNA gene polymerase chain reaction and pyrosequencing.
  
  
  
  We isolated and identified 101 distinct bacterial strains spanning 6 phyla including (1) novel strains with <98% 16S rRNA sequence identity to validly described species, (2) closely related species within a genus, (3) bacteria previously isolated from body sites other than the vagina, and (4) known bacteria formerly isolated from the vagina. Pyrosequencing showed that novel strains Peptoniphilaceae DNF01163 and Prevotellaceae DNF00733 were prevalent in women with BV.
  
  
  
  We isolated a diverse set of novel and clinically significant anaerobes from the human vagina using conventional approaches with systematic molecular identification. Several previously "uncultivated" bacteria are amenable to conventional cultivation.
authors.affiliation: Vaccine and Infectious Disease Division, Fred Hutchinson Cancer Research Center.
authors.forename: Sujatha
authors.initials: S
authors.lastname: Srinivasan
authors.affiliation: Vaccine and Infectious Disease Division, Fred Hutchinson Cancer Research Center.
authors.forename: Matthew M
authors.initials: MM
authors.lastname: Munch
authors.affiliation: Department of Biology, Northeastern University, Boston, Massachusetts.
authors.forename: Maria V
authors.initials: MV
authors.lastname: Sizova
authors.affiliation: Vaccine and Infectious Disease Division, Fred Hutchinson Cancer Research Center.
authors.forename: Tina L
authors.initials: TL
authors.lastname: Fiedler
authors.affiliation: Vaccine and Infectious Disease Division, Fred Hutchinson Cancer Research Center.
authors.forename: Christina M
authors.initials: CM
authors.lastname: Kohler
authors.affiliation: Department of Laboratory Medicine.
authors.forename: Noah G
authors.initials: NG
authors.lastname: Hoffman
authors.affiliation: Vaccine and Infectious Disease Division, Fred Hutchinson Cancer Research Center.
authors.forename: Congzhou
authors.initials: C
authors.lastname: Liu
authors.affiliation: Department of Obstetrics and Gynecology.
authors.forename: Kathy J
authors.initials: KJ
authors.lastname: Agnew
authors.affiliation: Department of Medicine, University of Alabama, Birmingham.
authors.forename: Jeanne M
authors.initials: JM
authors.lastname: Marrazzo
authors.affiliation: Department of Biology, Northeastern University, Boston, Massachusetts.
authors.forename: Slava S
authors.initials: SS
authors.lastname: Epstein
authors.affiliation: Vaccine and Infectious Disease Division, Fred Hutchinson Cancer Research Center Department of Medicine Department of Microbiology, University of Washington, Seattle.
authors.forename: David N
authors.initials: DN
authors.lastname: Fredricks
mesh_terms: D001421:Bacteria, Anaerobic / Q000145:classification / Q000166:cytology / Q000235:genetics / Q000302:isolation & purification*; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D004275:DNA, Ribosomal / Q000737:chemistry / Q000235:genetics; D005260:Female; D006194:Gardnerella vaginalis / Q000145:classification / Q000166:cytology / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D064307:Microbiota; D016133:Polymerase Chain Reaction; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D017422:Sequence Analysis, DNA; D014621:Vagina / Q000382:microbiology; D016585:Vaginosis, Bacterial / Q000382:microbiology*
chemical_list: D004269:DNA, Bacterial; D004275:DNA, Ribosomal; D012336:RNA, Ribosomal, 16S
keywords: anaerobes; bacterial vaginosis; cultivation; vagina