J Kikhney1, D von Schöning2, I Steding2, J Schulze3, A Petrich4, A Hiergeist5, U Reischl5, A Moter6, A Thomas2. 1. Biofilmzentrum, German Heart Institute Berlin, Berlin, Germany; Institut für Mikrobiologie und Hygiene, Charité - Universitätsmedizin Berlin, Campus Charité Benjamin Franklin, Berlin, Germany. 2. Klinik für Geburtsmedizin, Charité - Universitätsmedizin Berlin, Campus Charité Mitte, Berlin, Germany. 3. Biofilmzentrum, German Heart Institute Berlin, Berlin, Germany. 4. Institut für Mikrobiologie und Hygiene, Charité - Universitätsmedizin Berlin, Campus Charité Benjamin Franklin, Berlin, Germany. 5. Institute of Clinical Microbiology and Hygiene - University Hospital Regensburg, Regensburg, Germany. 6. Biofilmzentrum, German Heart Institute Berlin, Berlin, Germany. Electronic address: moter@dhzb.de.
Abstract
OBJECTIVES: Aim of this study was to detect microorganisms in fetal membranes and placental tissue in preterm chorioamnionitis by combining fluorescence in situ hybridization (FISH) with broad range PCR. The combination of the two molecular techniques enables identification and localization of the microorganisms within the tissue, confirming their clinical relevance. METHODS: In a prospective cohort study, we compared 31 women with preterm premature rupture of membranes or preterm labour and preterm delivery by caesarean section with a control group of 26 women undergoing elective caesarean section at term. Fetal membranes and placental tissue were analysed by FISH and broad range 16S rRNA-gene PCR and sequencing. RESULTS: For 20 women in the preterm group, caesarean section was performed because of a clinical diagnosis of chorioamnionitis. Microorganisms were detected in the tissues by both molecular techniques in 11 out of 20 women. Among those, Ureaplasma spp. was most abundant, with five cases that remained culture-negative and would have been missed by routine diagnostic procedures. Other infections were caused by Staphylococcus aureus, Streptococcus mitis or Escherichia coli. FISH and PCR were negative for all women without suspected chorioamnionitis and for the control group. CONCLUSIONS: Combination of FISH with broad-range PCR and sequencing permitted unambiguous identification of the causative microorganisms in chorioamnionitis. The high prevalence of Ureaplasma spp. should lead to a re-evaluation of its clinical significance and possible therapeutic consequences.
OBJECTIVES: Aim of this study was to detect microorganisms in fetal membranes and placental tissue in preterm chorioamnionitis by combining fluorescence in situ hybridization (FISH) with broad range PCR. The combination of the two molecular techniques enables identification and localization of the microorganisms within the tissue, confirming their clinical relevance. METHODS: In a prospective cohort study, we compared 31 women with preterm premature rupture of membranes or preterm labour and preterm delivery by caesarean section with a control group of 26 women undergoing elective caesarean section at term. Fetal membranes and placental tissue were analysed by FISH and broad range 16S rRNA-gene PCR and sequencing. RESULTS: For 20 women in the preterm group, caesarean section was performed because of a clinical diagnosis of chorioamnionitis. Microorganisms were detected in the tissues by both molecular techniques in 11 out of 20 women. Among those, Ureaplasma spp. was most abundant, with five cases that remained culture-negative and would have been missed by routine diagnostic procedures. Other infections were caused by Staphylococcus aureus, Streptococcus mitis or Escherichia coli. FISH and PCR were negative for all women without suspected chorioamnionitis and for the control group. CONCLUSIONS: Combination of FISH with broad-range PCR and sequencing permitted unambiguous identification of the causative microorganisms in chorioamnionitis. The high prevalence of Ureaplasma spp. should lead to a re-evaluation of its clinical significance and possible therapeutic consequences.
Authors: Gilles Faron; Ellen Vancutsem; Anne Naessens; Ronald Buyl; Leonardo Gucciardo; Walter Foulon Journal: Infect Dis Obstet Gynecol Date: 2017-02-05
Authors: Kenichiro Motomura; Roberto Romero; Yi Xu; Kevin R Theis; Jose Galaz; Andrew D Winters; Rebecca Slutsky; Valeria Garcia-Flores; Chengrui Zou; Dustyn Levenson; Robert Para; Madison M Ahmad; Derek Miller; Chaur-Dong Hsu; Nardhy Gomez-Lopez Journal: mBio Date: 2020-06-23 Impact factor: 7.867