Ureaplasma spp., commonly identified in the vagina/cervix of pregnant women with spontaneous preterm delivery, are the most frequently detected microorganisms in amniotic fluid. To date, 14 U. spp. serotypes have been characterized; however, modern molecular biology methods can distinguish different U. spp genotypes. Considering these factors, a knowledge gap exists regarding the association between U. spp. genotypes and the risk of the ascension of U. spp. from the cervix to the amniotic cavity. To fill this gap, an expanded multilocus sequence-typing scheme of U. spp. was performed to assess the relationship between cervical and amniotic fluid U. spp. in pregnant women with spontaneous preterm delivery. This study included 109 and 69 pregnant women with spontaneous preterm labor (PTL) and preterm prelabor rupture of membranes (PPROM), respectively. U. spp. DNA in cervical fluid was identified in 49% and 55% of the women with PTL and PPROM, respectively. The concurrent presence of U. spp. DNA in amniotic fluid was observed in 17% and 59% of the pregnant women with PTL and PPROM, respectively. Among pregnant women with PTL and PPROM, 38 expanded sequence types of cervical U. spp. were identified. No associations were observed between specific genotypes, subgroups, or clusters of cervical U. spp. and the presence of amniotic fluid U. spp. in pregnant women with spontaneous preterm delivery.

• Keywords
• Amniotic fluid, Genital mycoplasmas, Invasive sampling, Non-invasive sampling, Preterm birth,
• MeSH
• Cervix Uteri * microbiology   MeSH
• DNA, Bacterial genetics   MeSH
• Adult MeSH
• Genotype MeSH
• Pregnancy Complications, Infectious microbiology   MeSH
• Humans MeSH
• Young Adult MeSH
• Multilocus Sequence Typing MeSH
• Amniotic Fluid microbiology   MeSH
• Obstetric Labor, Premature microbiology   MeSH
• Fetal Membranes, Premature Rupture microbiology   MeSH
• Premature Birth * microbiology   MeSH
• Pregnancy MeSH
• Ureaplasma * genetics   isolation & purification   classification   MeSH
• Ureaplasma Infections * microbiology   MeSH
• Check Tag
• Adult MeSH
• Humans MeSH
• Young Adult MeSH
• Pregnancy MeSH
• Female MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• DNA, Bacterial MeSH

The main aim of this study was to determine expanded sequence types (eSTs) of Ureaplasma species (U. spp.). DNA isolated from the amniotic fluid of pregnancies complicated by preterm prelabor rupture of membranes (PPROM) using an expanded multilocus sequence typing scheme. Additionally, the study sought to examine whether phylogenetic subgroups of U. spp. DNA differ with respect to maternal demographic and clinical parameters and selected aspects of short-term neonatal morbidity. This retrospective cohort study was focused on singleton pregnancies complicated by PPROM occurring between the gestational ages of 24+0 and 36+6 weeks, where amniocentesis was conducted to assess the intra-amniotic environment and the presence of U. spp. DNA in the amniotic fluid samples was confirmed. The stored aliquots of U. spp. DNA were used to assess differences in nucleotide sequences in six U. spp. genes (ftsH, rpL22, valS, thrS,ureG, and mba-np1) using the eMLST scheme. The expanded multilocus sequence typing scheme was performed in 73 samples of U. spp. DNA isolated from pregnancies complicated by PPROM. In total, 33 different U. spp. DNA eSTs were revealed, 21 (#20, 233-244, 248-251, 253, 255, 259, and 262) of which were novel. The most frequently identified eST was #41, identified in 18% (13/73) of the aliquots. Based on their genetic relationships, the U. spp. DNA was divided into two clusters and four subgroups [cluster I (U. parvum): A, 43% (n = 31); B, 15% (n = 11); and C, 26% (n = 19); cluster II (U. urealyticum): 1; 16% (n = 12)]. Cluster II had a higher rate of polymicrobial findings than cluster I (58% vs 16%; p = 0.005), while subgroup A had the highest rate of concomitant Mycoplasma hominis in the amniotic fluid samples (66%; p = 0.04). In conclusion, Ureaplasma spp. DNA obtained from PPROM consisted of 33 different eSTs of U. spp. DNA. No differences in maternal and neonatal characteristics were found among the phylogenetical subgroups of U. spp. DNA, except for a higher rate of polymicrobial amniotic fluid findings in those with U. urealyticumand the concomitant presence of M. hominis in the amniotic fluid in those with the presence of U. parvum.

• Keywords
• Genital mycoplasma, Microbial invasion of the amniotic cavity, Molecular biology, Mollicutes, Morbidity, Neonates, Preterm delivery, Sequencing,
• MeSH
• DNA, Bacterial analysis   genetics   MeSH
• Adult MeSH
• Phylogeny MeSH
• Gestational Age MeSH
• Pregnancy Complications, Infectious microbiology   MeSH
• Humans MeSH
• Multilocus Sequence Typing * MeSH
• Amniotic Fluid * microbiology   MeSH
• Fetal Membranes, Premature Rupture * microbiology   MeSH
• Retrospective Studies MeSH
• Pregnancy MeSH
• Ureaplasma * genetics   isolation & purification   MeSH
• Ureaplasma Infections * microbiology   MeSH
• Check Tag
• Adult MeSH
• Humans MeSH
• Pregnancy MeSH
• Female MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• DNA, Bacterial MeSH