SOURCE:

Faculty: Health Sciences And Technology
Department: Medical Laboratory Science

CONTRIBUTORS:

Ezeanya, C. C.
Agbakoba, N. R.
Enweani, I. B.

ABSTRACT:

Mycoplasma genitalium is a sexually transmitted pathogen of public health concern in reproductive age women. Infections caused by this organism ultimately lead to serious sequelae such as chronic pelvic pain, ectopic pregnancy, infertility and increased risk for HIV transmission. The insidious nature of M. genitalium makes it unculturable, thus requiring the use of DNA amplification assays. In describing a novel microbe, the 16S rRNA gene sequence is crucial most especially for many unculturable bacteria. Therefore, the aim of this work was to characterize M. genitalium isolated from female subjects resident in Benin City, Nigeria. This study was both population and hospital based, cross-sectional study with 100 participants recruited from among female students of Edo State School of Health Technology and 30 women with gynaecological cases recruited at the Obstetrics and Gynaecology Clinic of Central Hospital, Benin City. The participants were aged 15-39 years and were recruited for the study using simple random sampling method. Ethical approval was obtained from the institutional Research Ethics Committee of Edo State Hospitals’ Management Board. Informed consent was also obtained from the participants. Duplicate samples of Endocervical Swabs (ECS) were collected from all the subjects. One set of the ECS were assayed microscopically using Wet Mount, Gram Stain and Giemsa Staining techniques. Evaluation of the second set of ECS for sexually transmitted pathogens (M. genitalium, M. hominis, U. urealyticum, U. parvum, T. vaginalis and C. trachomatis) was done using Multiplex Polymerase Chain Reaction with 6 specific primers following DNA extraction. All the samples were assayed for M. genitalium DNA using 16SrRNA Gene sequencing method with genus and specie- specific primers while Phylogenetic Analysis was done by Maximum Likelihood method using Tamura-Nei Model with bootstrap on Molecular Evolutionary Genetics Analysis (MEGA) version 7.0. Statistical analysis was done using SPSS version 20. Of the 130 subjects, 36 were positive for Mycoplasma species. 16SrRNA gene detection prior to sequencing was positive for Mycoplasma genus and negative for M.genitalium based on genus and specie - specific primers respectively. The prevalence of the different isolated Mycoplasma species were as follows: M. genitalium (0.8%), M. hominis (7.7%), M. penetrans (0.8%), U. urealyticum (3.1%) and U. parvum (1.5%); however, M. genitalium emerged only among the students. The accession numbers were generated for each consensus sequence following submission to GenBank. On the other hand, high number of sexual partners (≥4) (OR=0.18, P=0.012), co-infection with other sexually transmitted (ST) pathogens: T. vaginalis and C. trachomatis (OR=0.51, P=0.040) were strongly associated with Mycoplasma infection. Phylogenetic analysis of M. genitalium revealed no shared phylogenetic relationship with other species previously reported. Phylogenomics study of M. species among the subjects and reference strains revealed that all M. hominis from the subjects were found to be of a common ancestral clade and had significant association with two China strains: EU596508 (CV = 81%) and EU596509 (CV = 99%). Despite the relatively low prevalence of M. genitalium (0.8%); M. genitalium (MG238565) emerged with no shared phylogenetic relationship with others previously reported. This implies that the specie circulating in our environment is relatively unique. High number of sexual partners and co-infection with sexually transmitted pathogens presented as possible risk factors among female subjects. The significant association found between M. hominis strains in our study and those of China strains indicates indiscriminate spread of these genital mycoplasmas and when left undetected in the female reproductive tract have the potential of resulting to serious sequelae such as infertility. It is therefore paramount to monitor the spread of circulating genital pathogens among reproductive age women for better containment of infection and prevention of serious sequelae.