Detection Of Ureaplasma Research Articles

Molecular detection of bacteria, placental inflammation, and neonatal sepsis risk.

To identify bacteria in umbilical cord tissue and investigate the association with placental inflammation and neonatal sepsis risk score. Retrospective cohort study from 2017-2019. RNA was extracted from umbilical cord tissue and NanoString nCounter used to identify seven bacteria genera. Sepsis risk score was calculated using the Kaiser sepsis calculator. Placental histopathology was abstracted from medical records. Detection of bacterial RNA in the umbilical cord (n = 96/287) was associated with high-stage maternal and fetal acute placental inflammation (maternal 35.4% vs 22.5%, p = 0.03 and fetal 34.4% vs 19.4%, p < 0.01) and maternal vascular malperfusion (36.5% vs 23.0%, p = 0.02). Detection of Ureaplasma spp. was also associated with increased sepsis risk score (1.5/1000 [0.6, 8.6] vs 0.9/1000 [0.2, 2.9], p = 0.04). Umbilical cord bacterial pathogens are linked to fetal and maternal placental inflammation and maternal vascular malperfusion during gestation and associated with increased sepsis risk score in the neonate.

Prevalence of Mollicutes among men who have sex with men and transgender women aged 15 to 19 years in Salvador, North-eastern Brazil

BackgroundSome species of Mollicutes have been associated with different pathologies of the urogenital tract in humans, with a high prevalence among adult men who have sex with men (MSM) and transgender women (TGW). However, few studies have been performed to investigate its prevalence among adolescents. In this study, we estimated the initial prevalence of Mycoplasma genitalium (MG), Mycoplasma hominis (MH), Ureaplasma urealyticum (UU), and Ureaplasma parvum (UP); the rate of misdiagnosis at different anatomical sites; and the associated factors with positive tests for Mollicutes among MSM and TGW aged 15 to 19 years enrolled in the PrEP1519 study.MethodsPrEP-1519 is the first study to investigate the effectiveness of pre-exposure prophylaxis for human immunodeficiency virus among adolescent MSM and TGW aged 15 to 19 in Latin America. Oral, anal, and urethral swabs were taken from 246 adolescents upon enrolment in the study to detect MG, MH, UU, and UP by quantitative polymerase chain reaction (qPCR). Bivariate and multivariate analyses were conducted by Poisson regression and 95% confidence intervals (95% CI) were estimated.ResultsThe prevalence of Mollicutes was 32.1%. UU was the most prevalent species (20.7%), followed by MH (13.4%), MG (5.7%), and UP (3.2%); 67.3% of the positive samples would have been missed if only urethral samples had been taken. Receptive anal sex (prevalence ratio [PR] = 1.79; 95% CI = 1.07–3.01) and clinical suspicion of sexually transmitted infection (PR = 1.62; 95% CI = 1.01–2.61) were factors associated with the detection of Mollicutes in general. Group sex (PR = 1.98; 95% CI = 1.12–3.50) and receptive anal sex (PR = 2.36; 95% CI = 0.95–5.86) were associated with the detection of Mycoplasma spp. No sociodemographic, clinical, or behavioural variable was significantly associated with the detection of Ureaplasma spp.ConclusionsA high prevalence of Mollicutes was observed among adolescent MSM and TGW, especially at extragenital sites. Further research is required to understand the epidemiological profile of high-risk adolescents in different regions and contexts, and to investigate the pathogenesis of Mollicutes in the oral and anal mucosa before routine screening can be recommended in clinical practice.

MYCO WELL D-ONE detection of Ureaplasma spp. and Mycoplasma hominis in sexual health patients in Wales

The genital mycoplasmas are a unique group of inherently antibiotic-resistant sexually transmitted bacteria, often associated with non-gonococcal urethritis and bacterial vaginosis. The MYCO WELL D-ONE is a culture-based assay that aims to detect these organisms whilst concurrently screening them for antibiotic resistance. Urine and/or swabs from 856 informed and consented participants attending Welsh sexual health clinics were subjected to MYCO WELL D-ONE analysis, alongside qPCR and culture titration methodologies to determine sensitivity, specificity, PPV, NPV and accuracy. Resistance was confirmed by CLSI-compliant susceptibility testing and genetic mechanisms determined. The MYCO WELL D-ONE displayed a sensitivity and specificity of 91.98% and 96.44% for the detection of Ureaplasma spp., with sensitivity and specificity values of 78.23% and 98.84% for Mycoplasma hominis, compared with qPCR. Swabs harboured significantly greater bacterial loads than urine samples for both Ureaplasma spp. and M. hominis. Levofloxacin resistance rates, mediated by Ser83Leu mutation in ParC, for Ureaplasma spp. were 0.54%. Tetracycline resistance rates, mediated by tet(M), were 0.54% and 2% for Ureaplasma spp. and M. hominis, respectively; sequence analysis of tet(M)-positive Ureaplasma spp. and M. hominis strains isolated from a single individual confirmed separate resistance gene origins. The MYCO WELL D-ONE is a sensitive and specific assay for the detection of Ureaplasma spp. and M. hominis in genitourinary medicine samples, facilitating the accurate detection of these organisms within low-technology environments. While good for antibiotic resistance screening, accurate confirmation by MIC determination or molecular methods are required, and more optimally performed on urine samples.

Detection of ureaplasmas and bacterial vaginosis associated bacteria and their association with non-gonococcal urethritis in men.

No aetiology is found in up to 40% of men with symptomatic urethritis. Male partners of women with bacterial vaginosis (BV) may be at higher risk of non-gonococcal urethritis (NGU). The aim of this study was to examine the role of BV associated bacteria in first-void urine (FVU) in 97 asymptomatic men without urethritis (controls) and 44 men (cases) with NGU including 20 men with idiopathic urethritis (IU) attending a Swedish STD-clinic between January and October 2010. BV-associated bacteria and ureaplasmas were detected by quantitative PCR assays. All BV associated bacteria, except Megasphaera-like type 1, were strongly positively correlated with U. urealyticum p<0.005 and even stronger with the combined U. urealyticum and U. parvum load (p<0.0005) suggesting that ureaplasma induced elevated pH may stimulate the growth of BV associated bacteria. No statistically significant differences were found between IU cases and controls in the prevalence or load of BV associated bacteria or ureaplasmas. In multiple logistic regression, Megasphaera-like type 1 was associated with IU (p = 0.03), but most positive FVU samples contained very few bacteria and the finding may not be clinically relevant.

Mycoplasma Duo Test versus Conventional Culture Media for Detection of Ureaplasma in Endotracheal Aspirates from Respiratory Distressed Premature Neonates

Mycoplasma Duo Test versus Conventional Culture Media for Detection of Ureaplasma in Endotracheal Aspirates from Respiratory Distressed Premature Neonates

Detection of Ureaplasma spp. in semen samples from sheep in Brazil

A study was conducted to verify the presence of mycoplasmas and ureaplasmas DNA in sheep semen samples from the State of Pernambuco. The PCR assay was conducted of according with standard protocols with generic primers. Mollicutes DNA was detected in 26.0% and Ureaplasma spp. in 12.0% of semen samples.

Detection of Ureaplasma urealyticum and Ureaplasma parvum in amniotic fluid: association with pregnancy outcomes

Objective. Ureaplasma urealyticum is one of the organisms most frequently isolated from the amniotic fluid of women with adverse pregnancy. The prevalence of U. urealyticum and U. parvum on samples of amniotic fluid from healthy asymptomatic pregnant women, and whether its detection is associated with P-PROM or preterm birth was investigated.Methods. Transabdominal amniotic fluid obtained from 121 asymptomatic women at 16–20 weeks of gestation were tested for the detection of Ureaplasma spp., using a selective culture media. A Multiplex-Polymerase Chain Reaction (PCR) method was used for the identification of U. parvum and U. urealyticum. Pregnancy outcomes were obtained after the completion of all testing.Results. Ureaplasma spp. was not identified by culture, but was identified by Multiplex-PCR in four subjects, two corresponding to U. parvum and two to U. urealyticum. The women positive to ureaplasmas had normal labor, and babies born from infected-ureaplasmas pregnant women had normal weight birth. Preterm birth with intact membranes was documented in four women, all negative to ureaplasmas, but associated with gestational hypertension, lost of liquids and low weight birth.Conclusions. Multiplex-PCR method was more sensitive that culture in detecting ureaplasma organism in amniotic fluid. No association of ureaplasmas with pregnancy outcomes was found.

Detection of ureaplasmas by the polymerase chain reaction in the amniotic fluid of patients with cervical insufficiency

The purpose of this study was to determine the clinical significance of detecting microbial footprints of ureaplasmas in amniotic fluid (AF) using specific primers for the polymerase chain reaction (PCR) in patients presenting with cervical insufficiency. Amniocentesis was performed in 58 patients with acute cervical insufficiency (cervical dilatation, > or =1.5 cm) and intact membranes, and without regular contractions (gestational age, 16-29 weeks). AF was cultured for aerobic and anaerobic bacteria as well as genital mycoplasmas. Ureaplasmas (Ureaplasma urealyticum and Ureaplasma parvum) were detected by PCR using specific primers. Patients were divided into three groups according to the results of AF culture and PCR for ureaplasmas: those with a negative AF culture and a negative PCR (n=44), those with a negative AF culture and a positive PCR (n=10), and those with a positive AF culture regardless of PCR result (n=4). 1) Ureaplasmas were detected by PCR in 19.0% (11/58) of patients, by culture in 5.2% (3/58), and by culture and/or PCR in 22.4% (13/58); 2) Among the 11 patients with a positive PCR for ureaplasmas, the AF culture was negative in 91% (10/11); 3) Patients with a negative AF culture and a positive PCR for ureaplasmas had a significantly higher median AF matrix metalloproteinase-8 (MMP-8) concentration and white blood cell (WBC) count than those with a negative AF culture and a negative PCR (P<0.001 and P<0.05, respectively); 4) Patients with a positive PCR for ureaplasmas but a negative AF culture had a higher rate of spontaneous preterm birth within two weeks of amniocentesis than those with a negative AF culture and a negative PCR (P<0.05 after adjusting for gestational age at amnio-centesis); 5) Of the patients who delivered within two weeks of amniocentesis, those with a positive PCR for ureaplasmas and a negative AF culture had higher rates of histologic amnionitis and funisitis than those with a negative AF culture and a negative PCR (P<0.05 after adjusting for gestational age at amniocentesis, for each); 6) However, no significant differences in the intensity of the intra-amniotic inflammatory response and perinatal outcome were found between patients with a positive AF culture and those with a negative AF culture and a positive PCR. 1) Cultivation techniques for ureaplasmas did not detect most cases of intra-amniotic infection caused by these microorganisms (91% of cases with cervical insufficiency and microbial footprints for ureaplasmas in the amniotic cavity had a negative AF culture); 2) Patients with a negative AF culture and a positive PCR assay were at risk for intra-amniotic and fetal inflammation as well as spontaneous preterm birth.

Molecular detection ofUreaplasma urealyticum infection from clinical urogenital swabs

A simple nucleic acid amplification test (NAAT) was developed for detection of Ureaplasma urealyticum infection based on the PCR amplification of the urease gene (UU1/UU2 Test). DNA was extracted from urogenital swabs and a 225-bp long DNA fragment was amplified by PCR. NAAT was compared to the commercial amplification kit for sexually transmitted disease reference assay. The sensitivity and specificity of the UU1/UU2 Test were determined to be 100 and 98.9%, respectively. The overall prevalence rate in this group of patients was found to be about 236 per 1000 (283 and 166 per 1000 in females and males, respectively). These data demonstrate that UU1/UU2 Test is suitable for effective epidemiological screening and/or diagnostic practice.

Detection of Ureaplasma DNA in Endotracheal Samples Is Associated With Bronchopulmonary Dysplasia After Adjustment for Multiple Risk Factors

Microorganisms are hypothesized to contribute to the pathogenesis of bronchopulmonary dysplasia (BPD) in very low birth weight (VLBW) infants. This hypothesis remains controversial. We sought to determine whether endotracheal colonization with Ureaplasma sp., adenovirus, or Chlamydia sp. increases the risk of BPD. Intubated VLBW infants were included. Polymerase chain reaction (PCR) analysis was used to detect Ureaplasma sp., adenovirus, and Chlamydia sp. The outcome measure was BPD or death due to lung disease. Detection of microorganisms was compared between subjects with and without BPD. Logistic regression was used to control for covariates. Of 139 subjects, 33 (25%) screened positive for Ureaplasma sp., 22 of 136 (16%) were positive for adenovirus; eight of 133 (6%) were positive for Chlamydia sp. At 36 wk postmenstrual age, 14 patients had died, 68 (57%) had BPD. Detection of Ureaplasma sp. was associated with BPD or death (p < 0.001); adenovirus (p = 0.52) and Chlamydia sp. (p = 0.33) were not. Controlling confounding factors, the odds ratio for Ureaplasma sp. and BPD or death was 4.2 (95% CI 1.03, 17). In our population, detection of Ureaplasma sp., but not adenovirus or Chlamydia sp. was associated with BPD or death due to lung disease.

Value of Endocervical Ureaplasma Species Colonization as a Marker of Preterm Delivery

Aims: To study the predictive value for preterm delivery of colonization of the cervix and vagina by ureaplasmas and other potentially pathogenic microorganisms. Methods: Prospective analysis of a study group of 200 pregnant women with preterm labor and intact membranes, and a control group of 50 pregnant women. The subjects in both groups were between 24 and 34 weeks of gestation. Ureaplasma spp. and Mycoplasma hominis endocervical cultures were performed for both groups. Results: In the study group, 70 of the women delivered preterm, and all women in the control group carried their pregnancies to full term. Cervical cultures for Ureaplasma spp. were positive in 119 women, and were isolated more frequently in the study group (51.5%) than in the control group (32%), and more frequently in subjects in the study group who delivered preterm (65%) than in those who had a full-term delivery (45%), differences that were statistically significant. Conclusion: Detection of Ureaplasma spp. in the endocervix has a statistically significant relationship to preterm labor (p = 0.03) and preterm delivery (p = 0.02) in pregnant women with preterm labor and intact membranes.

Antibodies to Chlamydia trachomatis heat-shock protein 60 kDa and detection of Mycoplasma genitalium and ureaplasma urealyticum are associated independently with chronic nongonococcal urethritis.

Chronic nongonococcal urethritis (NGU) is a well-recognized clinical problem in genitourinary medicine clinics, but its etiology and optimal management are poorly understood. The authors showed previously that antibody to chlamydial hsp60 is associated with urethritis 30 to 92 days after treatment of acute NGU (chronic NGU) and that the detection of ureaplasmas or is associated with chronic NGU in which symptoms or signs are present. The aim was to determine whether these associations are independent of each other. This was a longitudinal prospective follow-up study over a 3-month period of 86 men with acute NGU. Men were tested for, ureaplasmas, and antibody to chlamydia hsp60 at presentation and during follow-up. The detection of either ureaplasmas or (OR, 29.45; 95% CI, 1.78-487) and the occurrence of hsp60 antibody at 30 to 92 days' follow-up (OR, 26.45; 95% CI, 1.34-523) were associated independently with the development of chronic NGU 30 to 92 days after treatment of acute NGU. The presence of chlamydial hsp60 antibody at 30 to 92 days was not associated with the development of chronic NGU in which symptoms or signs were present (P= 0.363). Chlamydial hsp60 antibody associated, however, with chronic NGU in which there were no symptoms or signs (P = 0.01). The results suggest that the immune response to chlamydial hsp60 may have a role in the etiology of chronic NGU in asymptomatic men who have no discharge on examination. The clinical relevance of this is unknown.

Do all men attending departments of genitourinary medicine need to be screened for non-gonococcal urethritis?

We investigated the influence of symptoms and signs on the detection of Chlamydia trachomatis, Mycoplasma genitalium and Ureaplasma urealyticum organisms (ureaplasmas) in men with non-gonococcal urethritis (NGU). Two hundred and forty-two men attending the Jefferiss Wing at St Mary's Hospital for a sexual health assessment were evaluated, of whom 169 had NGU. Urethral inflammation was diagnosed if there were either > or =5 polymorphonuclear leucocytes (PMNLs) per high-power field (HPF) in five or more microscope fields of a Gram-stained urethral smear, or > or =10 PMNLs per HPF in five or more fields of a Gram-stained thread from 15-20 mL of a first-passed urine (FPU) specimen. C. trachomatis was diagnosed by direct immunofluoresence, M. genitalium by a polymerase chain reaction assay and ureaplasmas by culture. On multivariate analysis, to control for potential confounding by age, ethnicity, sexual lifestyle and co-infection, an urethral discharge remained significantly associated with the detection of C. trachomatis and M. genitalium in men with acute urethritis [OR 12.3, 95% CI (2.39-63.5) and OR 35.2, 95% CI (3.9-319.6), respectively], but dysuria or penile irritation did not. The detection of ureaplasmas was not associated with any clinical feature. In addition, on multivariate analysis men with NGU who were either symptomatic or had an observable discharge were more likely to have C. trachomatis or M. genitalium detected [(OR 6.92, 95% CI 1.41-33.9) and (OR 5.18, 95% CI 0.99-27.1), respectively], but not ureaplasmas (OR 1.19, 95% CI 0.33-4.35). The findings suggest that in men with acute NGU, symptoms or signs, and in particular a urethral discharge, are associated with the detection of C. trachomatis and M. genitalium, but not ureaplasmas. Currently, there is no precise answer to the question of whether all men attending a GUM clinic need to be screened for NGU, but if clinically asymptomatic NGU is found not to be associated with a sexually transmitted pathogen, the UK clinical guidelines requiring the preparation of a urethral smear from such men would need to be revised.

Detection of Ureaplasma urealyticum in endotracheal tube aspirates from neonates by PCR.

A PCR-based test was optimized for the detection of Ureaplasma urealyticum from neonatal respiratory specimens, with primers directed against the multiple-banded antigen gene (L. J. Teng, X. Zheng, J. I. Glass, H. Watson, J. Tsai, and G. H. Cassell, J. Clin. Microbiol. 32:1464-1469, 1994). Endotracheal tube aspirates (225) from 103 low-birth-weight neonates (<1,250 g) were taken, when possible, at days 0, 4, and 14 after birth and examined by culture and by PCR. Of 77 specimens positive by either method, 73 were detected by PCR and 60 were detected by culture. Overall, 36% of the neonates were positive for U. urealyticum by either method. Of 16 patients with PCR-positive-culture-negative results, 13 had positive cultures at another sampling point, and one additional patient had a twin with positive cultures. Of 11 patients with day 0 specimens positive by PCR alone, 9 subsequently became culture positive, demonstrating the utility of this test in early detection. Multiple serovars were present in over 50% of positive specimens, with serovars 3 and 14 in combination being most prevalent. The amplicon size generated from the specimen by PCR correctly predicted the biovars isolated in over 85% of positive specimens. Thus, this PCR test was valuable in allowing early detection of U. urealyticum in neonatal respiratory specimens, as well as in providing biovar information.