Detection Of Ureaplasma Urealyticum Research Articles

Cloning, expression and purification of antigenic fragments of the Ureaplasma urealyticum UreD protein and its value in serology.

The detection of Ureaplasma urealyticum is usually done through culture. With the change of the smallest effective factor in culture, we face the lack of growth of these bacteria, which is one of the important reasons to find a suitable alternative for the diagnosis of this bacterium. UreD is a protected gene in this bacterium. The aim of this was to evaluate the ability of antigenic regions of UreD protein to bind to patients' serum antibodies. Antigenic regions of UreD protein were predicted using IEDB software with five different methods: Emini Surface Accessibility Prediction, Kolaskar and Tongaonkar Antigenicity, Chou and Fasman beta turn prediction, Karplus and Schulz flexibility scale, Ellipro-Epitope prediction based upon structural protrusion. Antigenic regions of UreD gene was clonned, expressed and purified. The antigenicity of this recombinant protein against the antibodies in the serum of people infected with U. urealyticum infections was checked in western blotting. The results showed that the antigenic regions of the UreD protein was producted and its antigenicity was demonstrated in western blotting. Moreover, all sera from patients infected with U. urealyticum reacted to the recombinant antigen. Specimens from people infected with U. urealyticum infection was positive in Western blotting suggesting that UreD protein has antigenic properties. Therefore, it can be used as a suitable candidate for the design of diagnostic kits and U. urealyticum vaccine.

Evaluation of the real-time fluorescence loop-mediated isothermal amplification assay for the detection of Ureaplasma urealyticum

Ureaplasma urealyticum (UU) is commonly present in human reproductive tract, which frequently leads to genital tract infection. Hence, there is an urgent need to develop a rapid detection method for UU. In our study, a real-time fluorescence loop-mediated isothermal amplification (LAMP) assay was developed and evaluated for the detection of UU. Two primers were specifically designed based on the highly conserved regions of ureaseB genes. The reaction was carried out for 60 min in a constant temperature system using Bst DNA polymerase, and the process was monitored by real-time fluorescence signal, while polymerase chain reaction (PCR) was performed simultaneously. In real-time fluorescence LAMP reaction system, positive result was only obtained for UU among 9 bacterial strains, with detection sensitivity of 42 pg/μL (4.2 × 105 CFU/mL), and all 16 clinical samples of UU could be detected. In conclusion, real-time fluorescence LAMP is a simple, sensitive, specific and effective method compared with conventional PCR, which shows great promise in the rapid detection of UU.

Detection of ureaplasma urealyticum by polymerase chain reaction examination in nonspecific genital infection patients

Non specific genital infection (NSGI) is a condition affecting females which causes inflammation of the endocervix or anterior urethra that is not caused by Neisseria gonorrhoeae. The causative sexually transmitted organisms include Chlamydia trachomatis (Groups D to K) and Ureaplasma urealyticum. Infection caused by Ureaplasma urealyticum is often asymptomatic even though many studies have pronounced that Ureaplasma urealyticum can contribute not only to lower genitourinary infection but also to infertility. Ureaplasma urealyticum cannot be stained by Gram stain due to the lack of a cell wall of the organism. This research aims to evaluate the prevalence of Ureaplasma urealyticum in NSGI patients by using the polymerase chain reaction (PCR) method targeted in the ureaplasma gene structure 429 bp area. The samples were extracted from eighteen DNA NSGI patients. Eleven out of eighteen (61.11%) DNA NSGI samples tested positive for Ureaplasma urealyticum. Most patients (44.44%) with Ureaplasma urealyticum were unemployed, and 27.78% were complaining of recurrent vaginal discharge. The high incidence of Ureaplasma urealyticum in this study needs further attention since doxycycline remains the drug of choice of NSGI. Moxifloxacin should be considered for patients who are making no clinical progress with doxycycline.

Prediction of chorioamnionitis in cases of intraamniotic infection by ureaplasma urealyticum in women with very preterm premature rupture of membranes or preterm labour

Objectives: First, to determinate the frequency of chorioamnionitis and funisitis in cases of intramniotic detection of Ureaplasma urealyticum. Second, to assess the predictive capability of some biological markers in the amniotic fluid of these women to predict histological inflammation.Subjects and methods: We prospectively studied 20 cases of women with premature rupture of membranes or preterm labour (PROM) or preterm labour and intraamniotic detection of Ureaplasma urealyticum. Gestational age at admission was 26.74 ± 2.53 weeks. Amniotic fluid concentrations of IL18, IL 2, IL4, IL6, IL10, IL12, TNF-alpha, IFN-g, and MMP-8 were measured by the Multiplex method. Amniotic fluid glucose and leukocyte count were also measured by standard methods. Placental detailed histological studies were performed. Student’s t-test, forward stepwise conditional binary logistic regression analysis and ROC curves were used.Results: Histological chorioamnionitis was present in 45% of cases (9/20) and funisitis just in 15% (3/20). Interleukins 6, 8, 12, MMP-8, and leukocyte count were significantly elevated in cases of histological inflammation, defined as choriamnionitis or chorioamniotis + funisitis (p = .007, .03, .01, .03, .03, respectively) while glucose was decreased (p = .04). Binary logistic regression for the prediction of inflammation showed a high predictive value (R2 = .66, p = .002) including in the equation only the IL6 value.Conclusions: A significant percentage of cases with intraamniotic detection of Ureaplasma urealyticum shows no pathological signs of histological inflammation. Concentration of Interleukin 6 in amniotic fluid can be useful for the diagnosis of subclinical chorioamnionitis in these cases.

Patients with cervical Ureaplasma Urealyticum and Chlamydia Trachomatis infection undergoing IVF/ICSI-ET: The need for new paradigm.

Genital tract infections with ureaplasma urealyticum (UU) and chlamydia trachomatis (CT) are the most frequent sexually-transmitted disease worldwide. UU and CT infections are considered to be the leading cause for infertility and adverse pregnancy outcomes. However, little is known about the specific effect of cervical UU and CT infections on the etiology of female infertility, as well as the pregnancy outcomes of the patients undergoing in vitro fertilization/intracytoplasmic sperm injection-embryo transfer (IVF/ICSI-ET). In order to find the association between cervical UU and/or CT infection and pregnancy outcomes, we conducted a retrospective case-control study on the patients undergoing IVF/ICSI-ET with cervical UU and/or CT infection. A total of 2208 patients who received IVF/ICSI-ET were enrolled in this study. Data on the general conditions, pregnancy history and clinical pregnant outcomes were analyzed in terms of the cervical UU and CT detection. Our results revealed that cervical UU and CT infections were the risk factors for ectopic pregnancy and tubal factor-induced infertility. Moreover, the pregnancy rate, abortion rate, ectopic pregnancy rate and premature birth rate in patients with UU and/or CT infections showed no significant difference when compared with the control group. We recommend that cervical UU and CT detection should be an optional item for infertility patients and clinical UU detection should differentiate the subtypes of cervical UU. Positive cervical UU and CT infections should not be taken as strict contraindications for IVF/ICSI-ET.

Molecular Detection of Ureaplasma Urealyticum and Ureaplasma Parvum Colonization in Preterm Infants with Respiratory Distress Syndrome

Background: Early neonatal acquisition of Ureaplasma urealyticum (U. urealyticum) and Ureaplasma parvum (U. parvum), in the 1 st 24 hours after birth and during passage of an infected maternal birth canal, have been linked to the development of respiratory diseases in premature newborns as a result of respiratory tract colonization. However, the role of these pathogens in the development of respiratory distress syndrome (RDS) still needs more clarification. Objectives: In this study we aimed to search for U. urealyticum and U. parvum in tracheal aspirates of premature infants with RDS by species-specific PCR assays and to evaluate the short term outcome of preterm infants with positive results. Methodology: Tracheal aspirate fluid samples from 35 preterm neonates with RDS with a mean gestational age of (32.76±2.48) weeks were collected within the first 24 hours after birth and analyzed for detection of U. urealyticum and U. parvum by species-specific PCR assays targeting urease gene subunits and adjoining spacer regions. Results: Ureaplasma spp. was detected in the tracheal aspirate of 6 (17%) preterm infants with RDS. Among these 6 positive cases; 5 (83%) were positive for U. parvum while only 1 case (17%) was positive for U. urealyticum. The preterm infants with positive results for Ureaplasma spp. had statistically significant higher degree of RD depending on the chest X ray grading results and longer durations of ventilation and hospitalization. Conclusions: Limited by the relatively small sample size, this work can be considered as a pilot study indicating a possible association between the severity of RDS in preterm infants and respiratory tract (RT) colonization with Ureaplasma spp. particularly U.parvum.

What is Missing in STD Screening in Hong Kong?

This retrospective analysis was to study the prevalence of Sexually Transmitted Diseases (STD) in a Reproductive Medical Center in Hong Kong. A total of 1190 patients were included in this study. Group 1A, couples had no symptoms but presented with subinfertility; Group 1B, the subfertile couples were positive for either Chlamydia trachomatis (CT) or Ureaplasma urealyticum (UU); Group 2, couples with symptoms were offered full STD screening including CT, UU, Mycoplasma hominis (MH), Neisseria gonorrhoea (NG), Syphilis, Herpes Simplex Virus 1 and 2. The methods of ELISA and quantitative real-time PCR were used for these analyses. Group 1A: UU detection rates in both male and female (13.84 v.s. 37.07%) were significantly higher than that of CT (5.65 v.s. 5.57%); Group 1B and Group 2: For those who had Full STD check, UU and MH detection rates were significantly higher than that of CT (35, 13.7 and 7.1% respectively). Over 47% of patients showed positive for one or more organism. In the subfertile couples, the UU and CT detection rates were much higher in females than those in males. Both semen and urine samples gave the same rates of infection among CT, UU, NG and MH. U. urealyticum infection rate rather than Chlamydis trachomatis infection is highest in Hong Kong. The infection rate in females is higher than in males. The detection rates in semen and urine samples in males are similar.

Resultados perinatales en amenazas de parto prematuro con colonización endocervical por Ureaplasma urealyticum

Objective To assess the perinatal results in pregnant women with threatened preterm labor and detection of Ureaplasma urealyticum by endocervical culture. Material and methods Seventy-two pregnant women with at least one episode of preterm labor between 24 and 36.6 weeks of pregnancy from January 2002 to December 2003 were included in our study. An endocervical culture for genital mycoplasmas was performed at admission. Exclusion criteria consisted of multiple pregnancy and premature rupture of membranes prior to the episode of threatened preterm labor. Perinatal results were compared in women with positive and negative cultures to U. urealyticum. Results There were 30 women with a U. urealyticum-positive culture and 42 women with a U. urealyticum-negative culture. There were no statistically significant differences in maternal age, nulliparity rate, cervical length or Bishop's score. Gestational age at delivery and perinatal results were highly similar in the two groups. There were no cases of chorioamnionitis or neonatal sepsis. Discussion In women admitted to hospital for threatened preterm labor, detection of U. urealyticum in endocervical culture at admission is not related to an increased risk of preterm birth or with increased perinatal morbidity.

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.

Prevalence and Detection of Ureaplasma urealyticum and Mycoplasma hominis in Endocervical Specimens from Women with Genitourinary Tract Diseases in Iran

Prevalence and Detection of Ureaplasma urealyticum and Mycoplasma hominis in Endocervical Specimens from Women with Genitourinary Tract Diseases in Iran

Detection of Ureaplasma urealyticum in Semen of Infertile Men by PCR

Ureaplasma urealyticum is a causative agent ofnon-gonococcal urethritis, prostatitis, epididymitis and infertility. The organism is more common in partners of infertile than fertile marriages. U. urealyticum infections not only jeopardize fertility but also pose a risk for infertility treatment and resulting pregnancies. The purpose of this study was to determine the prevalence of U. urealyticum in semen of infertile and healthy men by Polymerase Chain Reaction (PCR). Semen samples were obtained from infertile patients and healthy control and were subjected to the routine andrological analysis and PCR. DNA was extracted by Cadieux method and analyzed by PCR protocol with species-specific primers for U. urealyticum (urease gene). U. urealyticum was detected significantly by PCR in 12 of 100 (12%) semen specimens from infertile patients and in 3 of 100 (3%) healthy men. The volume of semen fluid, concentration of sperm cells and sperm cell with normal morphology were significantly decreased in infertile men. In the group of infertile patients with PCR positive for U. urealyticum the volume, count and morphology of semen samples were lower than in the infertile patients with PCR negative results.

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.

Perfil microbiológico en secreciones genitales de embarazadas sintomáticas, en el Gran Buenos Aires, Argentina

Establish the prevalence of microorganisms associated with genital colonization in symptomatic pregnant women. In order to review the evolution of frequent pathogens ecology and adjust the laboratory design, in a population attended at the public health Hospital, in the Great Buenos Aires. Vaginal and endocervical samples, were explored for specific detection of Chlamydia trachomatis, Neisseria gonorrhoeae, Streptococcus agalactie, Trichomonas vaginalis, Candida spp., Mycoplasma hominis, Ureaplasma urealyticum and bacterial vaginosis (VB).Direct methods, culture, inmunodetection and PCR, were employed. In 1999; 198 women, with genital discharge, were studied. Age in the group range from 16 to 42 years old (Median 27 years old). In 51 cases (25.7%) none of the above microorganisms or bacterial vaginosis were detected. In 30 cases (15.1%) bacterial vaginosis was diagnosed. Frequency of detection was: Ureaplasma urealyticum, 49,5%; Candida spp., 34,3%; Mycoplasma hominis, 14.1%; Streptococcus agalactie, 4.5%; Trichomonas vaginalis, 3.5%; Chlamydia trachomatis, 2.5%. No detection of Neisseria gonorrhoeae was demonstrated. There is a relevant frequency of bacterial vaginosis. On the other hand, lower prevalence of the Trichomonas vaginalis and Chlamydia trachomatis and also the absence of Neisseria gonorrhoeae was demonstrated. Culture for Streptococcus agalactie (at birth) and detection of Chlamydia trachomatis. must be extended to all pregnant women. Study of species and drug sensitivity of Candida spp., and detection of Ureaplasma urealyticum, Mycoplasma hominis and Neisseria gonorrhoeae, have to be explored under specific clinical requirement.

Detection of Ureaplasma urealyticum in urine of patients with systemic lupus erythematosus and healthy individuals by culture and polymerase chain reaction.

A method was developed to detect Ureaplasma urealyticum in urine by the polymerase chain reaction (PCR). A 457-bp fragment of the urease gene of U. urealyticum was amplified by PCR. Before PCR, components disturbing the amplification had to be reduced. This was possible by diluting the urine 1 in 10 with distilled water and by the extraction of the U. urealyticum DNA. Urine specimens from 41 patients with systemic lupus erythematosus (SLE) and 21 healthy individuals were treated by the dilution method and investigated by PCR for U. urealyticum DNA. The results were compared with those obtained by culture and the detection rates of PCR and culture were found to be identical. Also there was no difference in the detection rates of U. urealyticum from urine of SLE patients and healthy individuals; 10 (24.4%) of the 41 urine specimens from SLE patients and five (23.8%) of the 21 urine specimens from healthy individuals gave positive results for U. urealyticum. The results of this study do not indicate a decisive role for U. urealyticum in SLE.

Detection of Ureaplasma urealyticum in urethral swab samples from asymptomatic men and men with urethritis by a polymerase chain reaction-based assay

We compared a polymerase chain reaction (PCR)-based assay with primers specific for the 16S rRNA gene of Ureaplasma urealyticum with culture techniques for detecting U. urealyticum from urethral swab samples obtained from 256 asymptomatic men. Of 24 samples positive for U. urealyticum by culture, 23 samples were positive by the PCR-based assay, whereas 2 of 232 samples with a negative culture were positive by the PCR-based assay. The sensitivity and specificity for the PCR-based assay compared to culture were 95.8% and 99.1%, respectively. Our results confirmed the validity of the PCR-based assay for identifying this pathogen from urethral swab samples. In this study, we also examined urethral swab samples obtained from 195 men with urethritis for the detection of U. urealyticum by this assay. U. urealyticum was detected in 1 (3.4%) of 29 men with gonococcal urethritis and 23 (13.9%) of 166 men with nongonococcal urethritis. This assay may be a relevant tool to diagnose urethral Ureaplasma infections.

Detection of Ureaplasma urealyticum by polymerase chain reaction in the urogenital tract of adults, in amniotic fluid, and in the respiratory tract of newborns.

On the basis of the nucleotide sequence of the urease genes of Ureaplasma urealyticum serotype 8, polymerase chain reaction (PCR) primers were selected, evaluated for specificity and sensitivity, and tested for their ability to detect U. urealyticum in the adult urogenital tract, amniotic fluid, and endotracheal aspirates of newborns. All 14 reference serotypes of U. urealyticum were detected with equal sensitivity (1-10 cfu), whereas multiple strains of 12 other mycoplasma species found in humans as well as eukaryotic DNA were not detected. A total of 638 clinical specimens was evaluated. Results indicate that PCR is equal to if not more sensitive than culture for detection of U. urealyticum. Faster detection of U. urealyticum by PCR (< 24 h) compared to culture (2-5 days) will be particularly important in management of very low birth-weight infants in whom this organism has been shown to be a significant cause of meningitis, respiratory disease, and death. This method of detection will also be helpful in further determining the role of this organism in intraamniotic infection and premature birth.