Isolation and Molecular Identification of Mycoplasma Hominis in Infertile Female and Male Reproductive System (original) (raw)
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Scientific Reports
To our knowledge, the phylodistribution of M. hominis clinical strains associated with various pathological conditions of the urogenital tract has not been explored hitherto. Here we analyzed the genetic diversity and phylogenetic relationships among 59 M. hominis Tunisian clinical isolates, categorized as gynecological infections-or infertility-associated pathotypes. For this purpose, we developed an expanded multilocus sequence typing (eMLST) scheme, combining the previously reported multilocus sequence typing (MLST) loci (gyrB, tuf, ftsY, uvrA, gap) with a new selected set of putative virulence genes (p120', vaa, lmp1, lmp3, p60), referred herein to as multi-virulence-locus sequence typing (MVLST) loci. In doing so, M. hominis population was segregated into two distinct genetic lineages, which were differentially associated with each pathotype. Such a clear dichotomy was supported by several phylogenetic and population genetic analysis tools. Recombination was found to take place, but not sufficient enough to break down the overall clonal population structure of M. hominis, most likely as a result of purifying selection, which accommodated the most fit clones. In sum, and owing to the eMLST scheme described herein, we provide insightful data on the phylogenetics of M. hominis, arguing for the existence of genetically differentiable urogenital pathotypes. Mycoplasma hominis, which belongs to the Mycoplasmataceae family, in the Mollicutes class, was the first mycoplasma species isolated from humans in 1937 1. It resides, as a commensal, in the lower urogenital tract of healthy persons. Under certain circumstances, M. hominis can cause a variety of genital infections such as bacterial vaginosis, pelvic inflammatory disease, and cervicitis 2. This microorganism seems to be associated with pregnancy complications and neonatal diseases 3. In addition, several studies reported the pathogenic role of M. hominis in infertility 4,5. More interestingly, this species has been linked to a wide range of extragenital infections (septic arthritis, endocarditis, brain abscess), especially in immunocompromised patients 6-8. To better understand the epidemiology and the mode of spread of M. hominis, several molecular typing systems have been developed. These include Pulse-Field Gel Electrophoresis (PFGE), Restriction Fragment Length Polymorphism (RFLP) analysis, Amplified Fragment Length Polymorphism (AFLP), and Random Amplified Polymorphic DNA (RADP). All these methods have revealed a high degree of both genetic and antigenic heterogeneity among M. hominis strains 9-12. Although informative, these approaches proved to be quite difficult to
Iranian Journal of Reproductive Medicine, 2014
Background: Mycoplasmas can cause acute and chronic diseases at multiple sites with wide-range complications and have been implicated as cofactors in diseases. The infections influenced form genital mycoplasmas specifically Mycoplasma hominis and Mycoplasma genitalium potentially affect reproductive disorders, and infertility. Objective: Isolation and molecular identification of Mycoplasma genitalium from the genital tract of infertile male and vaginal discharge of infertile female referred to Infertility Center of Kerman in 2013. Materials and Methods: This study was a randomized, prospective study. We included 100 infertile male and 100 infertile female that were referred to the Infertility Center of Kerman. Then for isolation and molecular identification of Mycoplasma genitalium from urethral and vaginal discharge polymerase chain reaction was performed on Mycoplasma genus and genitalium. Results: From a total of 100 semen samples 45 patients (45%) were mycoplasma-positive and 13...
International Journal of Reproductive BioMedicine (IJRM)
Background: Genetic evidence of asymptomatic Mycoplasma hominis (M. hominis) and Ureaplasma urealyticum (U. urealyticum) infection associated with infertility among females is lacking because suitable high throughput molecular methods have not been applied. Objective: This study aimed to explore the occurrence of M. hominis and U. urealyticum in the genital tract of females with asymptomatic infection and infertility as well as determine their genetic relatedness. Materials and Methods: The study group included 100 asymptomatic females and 31 females diagnosed with infertility. Sequencing of the 16S rRNA gene following DNA extraction was performed directly from endo-cervical swabs. Phylogenetic analysis established the genetic linkage between the isolates from both groups. Results: In asymptomatic females, M. hominis and U. urealyticum were detected with a prevalence of 8% and 2% respectively. Among females with infertility, the prevalence was 6.45% and 3.23% for M. hominis and U. u...
Zentralblatt für Bakteriologie, 2000
Twenty Mycoplasma hominis strains isolated from colonized women and women with various urogenital infections were investigated for genetic and antigenic homo geneity by different methods. Restriction fragment length polymorphism analysis demonstrated heterogeneity for all strains, with one exception. Two strains sequen tially isolated from one patient showed identical patterns. Otherwise, no clonal clus tering could be detected within the strains isolated from either of the diagnostic groups. In contrast, SDS-PAGE analysis and the comparison of the immunoblot pat tern revealed antigenic similarities of strains isolated from patients with bacterial va ginosis, chorioamnionitis, premature rupture of membranes and preterm delivery as well as endometritis but showed obvious differences in comparison to strains isolat ed from colonized women.
Occurrence of Mycoplasma genitalium in fertile and infertile women
Fertility and Sterility, 2009
Objective: To determine the frequency of occurrence of Mycoplasma genitalium in the reproductive organs of infertile women in comparison with a control group of healthy, fertile women. Design: Prospective study. Setting: Gynecology Clinic at the 2nd Department of Gynecology and Obstetrics of the Wroclaw Medical University, Poland. Patient(s): The study included 51 patients with primary infertility (24 women with idiopathic infertility) and 23 women with proven fertility. Intervention(s): Cervical smear and smear from the peritoneal cavity, performed during laparoscopy. Main Outcome Measure(s): Presence of the genetic material of M. genitalium in the collected material analyzed using polymerase chain reaction (PCR). Result(s): M. genitalium was found in the cervical canal of 19.6% of all infertile patients and in 4.4% of fertile patients. In addition, the pathogen was discovered in the cervical canal of 29% patients with unexplained (idiopathic) infertility, which in comparison with the fertile group was a statistically significant difference. In the abdominal cavity, M. genitalium was found in 5.8% of patients from the infertile group (in 8.4% patients with idiopathic infertility), whereas it was not detected in the material obtained from the studied fertile patients.
Detection of Genital Mycoplasmas Infections among Infertile Females
Bahrain Medical Bulletin, 2013
the characteristic Ureaplasma and Mycoplasma hominis (M. hominis) colonies. Isolates were identified serologically as M. hominis or Ureaplasma urealyticum (U. urealyticum) by growth inhibition test (disc method). Result: Two genital mycoplasmas were detected out of 263 specimens by culture method. The positive specimens were identified as M. hominis. Conclusion: The result demonstrates lower values and further investigations for rapid detection of genital mycoplasmas in infertile female patients using PCR could be important and necessary for the detection of mycoplasmas infections.
Association of Mycoplasma genitalium with infertility in North Indian women
Indian journal of sexually transmitted diseases
Data regarding the association of Mycoplasma genitalium with infertility is scarce. This study was planned to look for the presence and association of M. genitalium in women with infertility. A prospective observational study was conducted on 100 cases of infertile women. The control group included 100 healthy fertile women. Samples of first void urine (FVU), endocervical swabs (ECS), and endometrial biopsies were subjected to polymerase chain reaction targeting MgPa gene to look for the presence of M. genitalium DNA. All endometrial biopsy samples were subjected to histopathological examination. A detailed clinical history of patients was taken, and all relevant investigations were recorded. M. genitalium was found in 16% of women with infertility from either of the samples that is, FVU and/or ECS and/or endometrium biopsy, and none from controls. ECS and biopsy could detect the highest number of cases (27%). Asymptomatic cases predominated in the study and M. genitalium positivity...
Research Journal of Microbiology, 2015
Mycoplasma hominis is normally found in the urinary tract of human and its role in Urinary Tract Infection (UTI) has been proved. This bacterium causes inflammatory responses and accumulation of leucocytes in urethra. In spite of the presence of the bacteria, the urine culture might be negative. Mycoplasma hominis can be transferred sexually and causes human infertility. The present study was conducted to detect and identify Mycoplasma hominis by molecular methods in urine samples of the patients with UTI, who were referred to our hospital. A total number of 864 urine samples from the patients with UTI were subjected to this study. After routine culture, urine analysis were performed on the samples. The DNA was extracted from the sediments of the urine samples, using phenol and chloroform method. Polymerase Chain Reaction (PCR) was conducted on the extracted DNA to detect the 16S rRNA of Mycoplasma hominis, with the primers; RNAH1 and RNAH2. Based on the results of PCR tests, out of 100 pyuria positive samples, 9 and 1% were infected with Mycoplasma sp. and Mycoplasma hominis, respectively. The sequencing of amplified product of 16S-rRNA revealed a single nucleotide substitution (269 T A), compared with the reference gene of this species.
Prevalence of Mycoplasma genitalium and Mycoplasma hominis in urogenital tract of Brazilian women
BMC Infectious Diseases, 2015
Background: The role of Mycoplasma hominis and M. genitalium in urogenital tract infections remains unknown. Furthermore these mollicutes present a complex relationship with the host immune response. The role of inflammatory cytokines in infections also makes them good candidates to investigate bacterial vaginosis and mycoplasma genital infections. Therefore, the aim of this study was to detect the above-mentioned mollicutes by quantitative Polymerase Chain Reaction (qPCR) methodologies in vaginal swabs and dosage of cytokines. Methods: Vaginal swabs and peripheral blood were collected from 302 women, including healthy individuals. The molecular findings were correlated with some individual behavioral variables, clinical and demographic characteristics, presence of other important microorganisms in vaginal swabs, and levels of interleukin (IL)-1β and IL-6. Results: M. hominis and M. genitalium were detected in 31.8% and 28.1% of samples, respectively. The qPCR results were associated with clinical signs and symptoms of the infections studied. The frequency of Trichomonas vaginalis, Gardnerella vaginalis, Neisseria gonorrhoeae and Chlamydia trachomatis was 3.0%, 21.5%, 42.4%, and 1.7% respectively. Increased levels of IL-1β were associated with the presence of M. hominis and signs and/or symptoms of the genital infection of women studied. Conclusion: IL-1β production was associated with the detection of M. hominis by qPCR. The sexual behavior of women studied was associated with the detection of mycoplasma and other agents of genital infections.