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

Carolyn Gardella; Donald E. Riley; Jane Hitti; Kathy Agnew; John N. Krieger; David Eschenbach

doi:10.1055/s-2004-831884

American Journal of Perinatology, Inhaltsverzeichnis

Am J Perinatol 2004; 21(6): 319-323
DOI: 10.1055/s-2004-831884

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

Abstract

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

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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