PDF herunterladen
Am J Perinatol 2004; 21(6): 319-323
DOI: 10.1055/s-2004-831884
Copyright © 2004 by Thieme Medical Publishers, Inc., 333 Seventh Avenue, New York, NY 10001, USA.

Identification and Sequencing of Bacterial rDNAs in Culture-Negative Amniotic Fluid from Women in Premature Labor

Carolyn Gardella1 , Donald E. Riley2 , Jane Hitti1 , Kathy Agnew1 , John N. Krieger1 , David Eschenbach1
  • 1Departments of Obstetrics and Gynecology, University of Washington, and VA Medical Center, Seattle, Washington
  • 2Department of Urology, University of Washington, and VA Medical Center, Seattle, Washington
Weitere Informationen

Publikationsverlauf

Publikationsdatum:
16. August 2004 (online)

Auch verfügbar auf
    Lizenzen und Reprints

Our objective was to identify bacterial species present in culture-negative but 16S rDNA-positive amniotic fluid samples from women in preterm labor. Amniotic fluid from 69 women in preterm labor was cultured and examined for the proinflammatory cytokine interleukin-6 (IL-6). Polymerase chain reaction technology was used to detect highly conserved bacterial ribosomal DNA sequences (16S rDNAs). As previously reported, 16S rDNAs were identified in 15 (94%) of 16 culture-positive amniotic fluid samples, in 5 (36%) of 14 culture-negative samples with elevated IL-6, and in 1 (3%) of 39 culture-negative samples with low IL-6 levels. Direct sequencing was performed of 16S rDNAs from the 5 culture-negative amniotic fluid specimens with elevated IL-6, followed by database searches and phylogenetic analyses. The bacterial sequences identified included: two Leptotrichia sanguinegens, one human oral bacterium A33, one Fusobacterium nucleatum, and one Ureaplasma urealyticum. Identification and sequencing of 16S rDNAs in amniotic fluid is a promising technique to identify bacterial species associated with elevated IL-6 levels in culture-negative amniotic fluid that may contribute to the etiology of premature labor.

KEYWORDS

16S rDNA - PCR - preterm labor - amniotic fluid - intra-amniotic infection

REFERENCES

  • 1 Hillier S L, Witkin S S, Krohn M A, Watts D H, Kiviat N B, Eschenbach D A. The relationship of amniotic fluid cytokines and preterm delivery, amniotic fluid infection, histologic chorioamnionitis, and chorioamnion infection.  Obstet Gynecol. 1993;  81 941-948
    PubMedSuche in Google Scholar
  • 2 Romero R, Yoon B H, Mazor M et al.. The diagnostic and prognostic value of amniotic fluid white blood cell count, glucose, interleukin-6 and gram stain in patients with preterm labor and intact membranes.  Am J Obstet Gynecol. 1993;  169 805-816
    PubMedSuche in Google Scholar
  • 3 Nikkari S, McLaughlin I J, Bi W, Dodge D E, Relman D A. Does blood of healthy subjects contain bacterial ribosomal DNA?.  J Clin Microbiol. 2001;  39 1956-1959
    CrossrefPubMedSuche in Google Scholar
  • 4 Relman D A, Schmidt T M, MacDermott R P, Fialkow S. Identification of the uncultured bacillus of Whipple’s disease.  N Engl J Med. 1992;  327 293-301
    CrossrefPubMedSuche in Google Scholar
  • 5 Palmer H M, Gilroy C B, Claydon E J, Taylor-Robinson D. Detection of Mycoplasma genitalium in the genitourinary tract of women by the polymerase chain reaction.  Int J STD AIDS. 1991;  2 261-263
    PubMedSuche in Google Scholar
  • 6 Relman D A. The search for unrecognized pathogens.  Science. 1999;  284 1308-1310
    CrossrefPubMedSuche in Google Scholar
  • 7 Hitti J, Riley D E, Krohn M A et al.. Broad-spectrum bacterial rDNA polymerase chain reaction assay for detecting amniotic fluid infection among women in preterm labor.  Clin Infect Dis. 1997;  24 1228-1232
    PubMedSuche in Google Scholar
  • 8 Relman D A, Falkow S. Identification of uncultured microorganisms: expanding the spectrum of characterized microbial pathogens.  Infect Agents Dis. 1992;  1 245-253
    PubMedSuche in Google Scholar
  • 9 Thompson J D, Higgins D G, Gibson T J. CLUSTAL W: Improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice.  Nucleic Acids Res. 1994;  22 4673-4680
    PubMedSuche in Google Scholar
  • 10 Hanff P A, Rosoldonoghue J, Spiegel C A, Wilson K H, Moore L H. Leptotrichia sanguinegens sp. Nov., a new agent of postpartum and neonatal bacteremia.  Clin Infect Dis. 1995;  20(suppl 2) S237-S239
    PubMedSuche in Google Scholar
  • 11 Collins M D, Hoyles L, Tornquvist E, vonEssen R, Falsen E. Characterization of some strains from human clinical sources which resemble “Leptotrichia sanguinegens”: Description of Sneathia sanguinegens sp. nov., gen.nov.  Syst Appl Microbiol. 2001;  24 358-361
    PubMedSuche in Google Scholar
  • 12 McGaw T. Periodontal disease and preterm delivery of low-birth-weight infants.  J Can Dent Assoc. 2002;  68 165-169
    PubMedSuche in Google Scholar
  • 13 Bearfield C, Davenport E S, Sivapathasundaram V, Allaker R P. Possible association between amniotic fluid micro-organism infection and microflora in the mouth.  BJOG. 2002;  109 527-533
    CrossrefPubMedSuche in Google Scholar
  • 14 Hitti J, Hillier S L, Agnew K J, Krohn M A, Riesner D P, Eschenbach D A. Vaginal indicators of amniotic fluid infection in preterm labor.  Obstet Gynecol. 2001;  97 211-219
    CrossrefPubMedSuche in Google Scholar
  • 15 Watts D H, Krohn M A, Hillier S L, Eschenbach D A. The association of occult amniotic fluid infection with gestational age and neonatal outcome among women in preterm labor.  Obstet Gynecol. 1992;  79 351-357
    CrossrefPubMedSuche in Google Scholar
  • 16 Gardella C, Hitti J, Martin T R, Ruzinski J, Eschenbach D A. Amniotic fluid lipopolysaccharide binding protein and soluble CD14 as mediators of the inflammatory response in preterm labor.  Am J Obstet Gynecol. 2001;  184 1241-1248
    CrossrefPubMedSuche in Google Scholar

Carolyn GardellaM.D. M.P.H. 

Department of Obstetrics and Gynecology, Box 356460, University of Washington Medical Center, Seattle, WA 98195; reprints not available from the author