PMID- 24302980
OWN - NLM
STAT- MEDLINE
DCOM- 20140625
LR  - 20211021
IS  - 1932-6203 (Electronic)
IS  - 1932-6203 (Linking)
VI  - 8
IP  - 10
DP  - 2013
TI  - More than meets the eye: associations of vaginal bacteria with gram stain 
      morphotypes using molecular phylogenetic analysis.
PG  - e78633
LID - 10.1371/journal.pone.0078633 [doi]
LID - e78633
AB  - Bacterial vaginosis (BV) is a highly prevalent condition associated with adverse 
      health outcomes. Gram stain analysis of vaginal fluid is the standard for 
      confirming the diagnosis of BV, wherein abundances of key bacterial morphotypes 
      are assessed. These Lactobacillus, Gardnerella, Bacteroides, and Mobiluncus 
      morphotypes were originally linked to particular bacterial species through 
      cultivation studies, but no studies have systematically investigated associations 
      between uncultivated bacteria detected by molecular methods and Gram stain 
      findings. In this study, 16S-rRNA PCR/pyrosequencing was used to examine 
      associations between vaginal bacteria and bacterial morphotypes in 220 women with 
      and without BV. Species-specific quantitative PCR (qPCR) and fluorescence in Situ 
      hybridization (FISH) methods were used to document concentrations of two bacteria 
      with curved rod morphologies: Mobiluncus and the fastidious BV-associated 
      bacterium-1 (BVAB1). Rank abundance of vaginal bacteria in samples with evidence 
      of curved gram-negative rods showed that BVAB1 was dominant (26.1%), while 
      Mobiluncus was rare (0.2% of sequence reads). BVAB1 sequence reads were 
      associated with Mobiluncus morphotypes (p<0.001). Among women with curved rods, 
      mean concentration of BVAB1 DNA was 2 log units greater than Mobiluncus (p<0.001) 
      using species-specific quantitative PCR. FISH analyses revealed that mean number 
      of BVAB1 cells was 2 log units greater than Mobiluncus cells in women with 
      highest Nugent score (p<0.001). Prevotella and Porphyromonas spp. were 
      significantly associated with the "Bacteroides morphotype," whereas Bacteroides 
      species were rare. Gram-negative rods designated Mobiluncus morphotypes on Gram 
      stain are more likely BVAB1. These findings provide a clearer picture of the 
      bacteria associated with morphotypes on vaginal Gram stain.
FAU - Srinivasan, Sujatha
AU  - Srinivasan S
AD  - Vaccine and Infectious Disease Division, Fred Hutchinson Cancer Research Center, 
      Seattle, Washington, United States of America.
FAU - Morgan, Martin T
AU  - Morgan MT
FAU - Liu, Congzhou
AU  - Liu C
FAU - Matsen, Frederick A
AU  - Matsen FA
FAU - Hoffman, Noah G
AU  - Hoffman NG
FAU - Fiedler, Tina L
AU  - Fiedler TL
FAU - Agnew, Kathy J
AU  - Agnew KJ
FAU - Marrazzo, Jeanne M
AU  - Marrazzo JM
FAU - Fredricks, David N
AU  - Fredricks DN
LA  - eng
GR  - R01 AI061628/AI/NIAID NIH HHS/United States
GR  - R01 HG005966/HG/NHGRI NIH HHS/United States
GR  - R01 HG005966-01/HG/NHGRI NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
DEP - 20131024
PL  - United States
TA  - PLoS One
JT  - PloS one
JID - 101285081
RN  - 0 (Gram's stain)
RN  - 0 (Phenazines)
RN  - 0 (RNA, Bacterial)
RN  - 0 (RNA, Ribosomal, 16S)
RN  - J4Z741D6O5 (Gentian Violet)
SB  - IM
MH  - Bacteroides/cytology/*genetics
MH  - Female
MH  - Gardnerella vaginalis/cytology/genetics
MH  - Gentian Violet
MH  - Humans
MH  - Lactobacillus/cytology/genetics
MH  - Mobiluncus/cytology/*genetics
MH  - Molecular Typing
MH  - Phenazines
MH  - Phylogeny
MH  - RNA, Bacterial/genetics
MH  - RNA, Ribosomal, 16S/genetics
MH  - Sequence Analysis, DNA
MH  - Vaginosis, Bacterial/*microbiology
PMC - PMC3840219
COIS- Competing Interests: The authors have declared that no competing interests exist.
EDAT- 2013/12/05 06:00
MHDA- 2014/06/26 06:00
PMCR- 2013/10/24
CRDT- 2013/12/05 06:00
PHST- 2013/08/29 00:00 [received]
PHST- 2013/09/04 00:00 [accepted]
PHST- 2013/12/05 06:00 [entrez]
PHST- 2013/12/05 06:00 [pubmed]
PHST- 2014/06/26 06:00 [medline]
PHST- 2013/10/24 00:00 [pmc-release]
AID - PONE-D-13-35638 [pii]
AID - 10.1371/journal.pone.0078633 [doi]
PST - epublish
SO  - PLoS One. 2013 Oct 24;8(10):e78633. doi: 10.1371/journal.pone.0078633. 
      eCollection 2013.