Detection and antimicrobial susceptibility of genital mycoplasmas in women and men, with 23S rRNA and parC mutation analysis in Mycoplasma genitalium.

Mycoplasma hominis and Mycoplasma genitalium are increasingly recognized for their potential association with reproductive disorders, yet regional data remain limited. This study simultaneously investigates infertile men and women in Hormozgan Province to determine genus- and species-level prevalence, assess susceptibility to key antibiotics, and identify resistance-associated mutations in M. genitalium.In this cross-sectional study, 400 participants (200 infertile and 200 fertile) were recruited from the Ome Leila Infertility Center, Bandar Abbas, Iran, during the first half of 2024. Fertility status was clinically assessed by specialist physicians. Inclusion criteria included no recent antibiotic use and sexual abstinence for 4-5 days prior to sample collection. Demographic and lifestyle data-including age, place of residence, smoking, and alcohol consumption-were collected via structured questionnaire. Endocervical swabs and semen samples underwent short-term enrichment in PPLO broth prior to genus- and species-specific PCR targeting the 16S rRNA gene. Minimum inhibitory concentrations (MICs) for erythromycin, azithromycin, and moxifloxacin were determined using the broth microdilution method on enriched cultures. Resistant M. genitalium isolates were sequenced for mutations in the 23S rRNA and parC genes. Binary logistic models were applied without adjustment for confounders; all analyses were considered exploratory.Mycoplasma DNA was detected in 22% of participants (n=88). Species-level identification revealed M. hominis in 9.8% (n=39) and M. genitalium in 12.3% (n=49), both more frequent among infertile individuals. MIC testing indicated macrolide resistance in 31.1% of M. genitalium isolates, with two also resistant to moxifloxacin. M. hominis exhibited lower resistance levels. Sequencing of resistant M. genitalium strains confirmed mutations in 23S rRNA (A2058G, A2059G) and parC (G248T). Among women, M. genitalium was more frequent in infertile participants, while M. hominis showed no significant association. In men, smoking and alcohol consumption were associated with infertility (OR=2.42; 95% CI: 1.21-4.87).This study provides region-specific molecular insights into genital Mycoplasma infections in southern Iran. While causal relationships cannot be established, the findings suggest that M. genitalium and M. hominis may be involved in infertility and support the potential value of including Mycoplasma screening in infertility evaluations alongside other infections.