Project description:Ureaplasma are widespread parasites colonizing the mucosal surface of the human urogenital tract, and it has been suspected as a causative agent of nongonococcal urethritis, pregnancy complications and prenatal infections. Ureaplasma may also cause central nervous system infections and affect the lower respiratory tract of newborn babies. However, Ureaplasma spp. have also been detected in the urogenital tracts of clinically healthy patients, and their role in the development of infections thus remains unclear. Like in other organisms, virulence of Ureaplasma is determined by the presence of virulence factors - adhesions, human IgA protease, phospholipase and urease. However, the existence of interrelationships between the presence of these genes in the Ureaplasma genome and the incidence of diseases in man has not been demonstrated. Difficulties in the elucidation of these interrelationships may arise from significant macro- (gene mutation, chromosomal rearrangements) and micro- (nucleotide polymorphism) genomic heterogeneity. It is possible that the combination of the variable strain-specific genes in Ureaplasma with generally known virulence factors determine the development of pathological processes on the mucosal surface of the human urogenital tract. Keywords: macroarray, variability of genome, Ureaplasma parvum
Project description:Ureaplasma parvum serovar 3 strain, OMC-P162, was isolated from the human placenta of a preterm delivery at 26 weeks' gestation. In this study, we sequenced the complete genome of OMC-P162 and compared it with other serovar 3 strains isolated from patients with different clinical conditions. Ten unique genes in OMC-P162, five of which encoded for hypothetical proteins, were identified. Of these, genes UPV_229 and UPV_230 formed an operon whose open reading frames were predicted to code for a DNA methyltransferase and a hypothetical protein, respectively. DNA modification analysis of the OMC-P162 genome identified N4-methylcytosine (m4C) and N6-methyladenine (m6A), but not 5-methylocytosine (m5C). UPV230 recombinant protein displayed endonuclease activity and recognized the CATG sequence, resulting in a blunt cut between A and T. This restriction enzyme activity was identical to that of the cultivated OMC-P162 strain, suggesting that this restriction enzyme was naturally expressed in OMC-P162. We designated this enzyme as UpaP162. Treatment of pT7Blue plasmid with recombinant protein UPV229 completely blocked UpaP162 restriction enzyme activity. These results suggest that the UPV_229 and UPV_230 genes act as a type II restriction-modification system in Ureaplasma OMC-P162.
Project description:Here, we present the complete genome sequence of Ureaplasma parvum serovar 3, clinical strain SV3F4, isolated from a Japanese patient with a history of an infectious abortion.
Project description:Ureaplasma are widespread parasites colonizing the mucosal surface of the human urogenital tract, and it has been suspected as a causative agent of nongonococcal urethritis, pregnancy complications and prenatal infections. Ureaplasma may also cause central nervous system infections and affect the lower respiratory tract of newborn babies. However, Ureaplasma spp. have also been detected in the urogenital tracts of clinically healthy patients, and their role in the development of infections thus remains unclear. Like in other organisms, virulence of Ureaplasma is determined by the presence of virulence factors - adhesions, human IgA protease, phospholipase and urease. However, the existence of interrelationships between the presence of these genes in the Ureaplasma genome and the incidence of diseases in man has not been demonstrated. Difficulties in the elucidation of these interrelationships may arise from significant macro- (gene mutation, chromosomal rearrangements) and micro- (nucleotide polymorphism) genomic heterogeneity. It is possible that the combination of the variable strain-specific genes in Ureaplasma with generally known virulence factors determine the development of pathological processes on the mucosal surface of the human urogenital tract. In our research we used 10 clinical and 1 laboratory strain
Project description:Objective. We compared laboratory developed real-time PCR assays for detection of Mycoplasma hominis and for detection and differentiation of Ureaplasma urealyticum and parvum to culture using genitourinary specimens submitted for M. hominis and Ureaplasma culture. Methods. 283 genitourinary specimens received in the clinical bacteriology laboratory for M. hominis and Ureaplasma species culture were evaluated. Nucleic acids were extracted using the Total Nucleic Acid Kit on the MagNA Pure 2.0. 5??L of the extracts were combined with 15??L of each of the two master mixes. Assays were performed on the LightCycler 480 II system. Culture was performed using routine methods. Results.??M. hominis PCR detected 38/42 M. hominis culture-positive specimens, as well as 2 that were culture negative (sensitivity, 90.5%; specificity, 99.2%). Ureaplasma PCR detected 139/144 Ureaplasma culture-positive specimens, as well as 9 that were culture negative (sensitivity, 96.5%; specificity, 93.6%). Of the specimens that tested positive for Ureaplasma species, U. urealyticum alone was detected in 33, U. parvum alone in 109, and both in 6. Conclusion. The described PCR assays are rapid alternatives to culture for detection of M. hominis and Ureaplasma species, and, unlike culture, the Ureaplasma assay easily distinguishes U. urealyticum from parvum.
Project description:BackgroundUreaplasma urealyticum (UUR) and Ureaplasma parvum (UPA) are sexually transmitted bacteria among humans implicated in a variety of disease states including but not limited to: nongonococcal urethritis, infertility, adverse pregnancy outcomes, chorioamnionitis, and bronchopulmonary dysplasia in neonates. There are 10 distinct serotypes of UUR and 4 of UPA. Efforts to determine whether difference in pathogenic potential exists at the ureaplasma serovar level have been hampered by limitations of antibody-based typing methods, multiple cross-reactions and poor discriminating capacity in clinical samples containing two or more serovars.ResultsWe determined the genome sequences of the American Type Culture Collection (ATCC) type strains of all UUR and UPA serovars as well as four clinical isolates of UUR for which we were not able to determine serovar designation. UPA serovars had 0.75-0.78 Mbp genomes and UUR serovars were 0.84-0.95 Mbp. The original classification of ureaplasma isolates into distinct serovars was largely based on differences in the major ureaplasma surface antigen called the multiple banded antigen (MBA) and reactions of human and animal sera to the organisms. Whole genome analysis of the 14 serovars and the 4 clinical isolates showed the mba gene was part of a large superfamily, which is a phase variable gene system, and that some serovars have identical sets of mba genes. Most of the differences among serovars are hypothetical genes, and in general the two species and 14 serovars are extremely similar at the genome level.ConclusionsComparative genome analysis suggests UUR is more capable of acquiring genes horizontally, which may contribute to its greater virulence for some conditions. The overwhelming evidence of extensive horizontal gene transfer among these organisms from our previous studies combined with our comparative analysis indicates that ureaplasmas exist as quasi-species rather than as stable serovars in their native environment. Therefore, differential pathogenicity and clinical outcome of a ureaplasmal infection is most likely not on the serovar level, but rather may be due to the presence or absence of potential pathogenicity factors in an individual ureaplasma clinical isolate and/or patient to patient differences in terms of autoimmunity and microbiome.