Introduction: Mycoplasma genitalium (MG) and Trichomonas vaginalis (TV) are Sexually Transmitted Infections (STIs) frequently detected in Non-chlamydial Non-gonococcal urethritis, however the prevalence of MG and TV in urine (U), anorectal (A) and pharyngeal (P) samples is unknown among men who have sex with men (MSM) in Belgium. The objective was to evaluate the MG/TV multiplex kit of Diagenode on the three different sample types.

Methods In the context of an ongoing study among MSM, U-A-P samples are collected every three months to test for STIs including MG and TV. DNA extraction is performed on the Abbott m2000sp system. The DNA extracts are tested in both an accredited in-house MG pdhD- gene real-time PCR (RT-PCR) for MG (target: pdhD gene) and the S-DiaMGTV multiplex kit of Diagenode for MG (target: mgpa and mg219 gene) and TV (target: TV-spec repeat gene). In addition, in case of a positive TV result, the sample is tested by an in-house TV RT-PCR (TV-spec repeat gene). All amplifications were performed using the Rotor-GeneQ MDx HRM (Qiagen) platform.

Results To date, of the 1779 samples that were tested for MG/TV, 1768 provided a valid result. Only one anorectal sample was positive for TV but this result could not be confirmed by the in-house TV RT-PCR assay. MG was detected by both assays in 64 samples. The S-DiaMGTV assay missed 3/2/1 U-A-P infections and identified additional 2/2/2 U-A-P infections. Both assays were 99.3% concordant for MG. Up to now an overall prevalence of 4.0% for MG is found: 4.8% (29/598); 6.4% (37/578) and 0.7% (4/592) for U-A-P, respectively. Compared to the in-house RT-PCR the S-DiaMGTV showed an overall sensitivity/specificity of 91.4%/99.7%: 89.7% (CI 95%: 72.7%–97.8%)/99.7% (CI 95%: 98.7%–100%) for urine; 94.6% (CI 95%: 81.8%–99.3%)/99.6% (CI 95%: 98.7%–100.0%) for anorectal and 75.0% (CI 95%: 19.4%–99.4%)/99.7% (CI 95%: 98.8%–100.0%) for pharyngeal samples.

Conclusion MG but not TV is prevalent in MSM in urine and anorectal samples. The S-DiaMGTV multiplex kit is an appropriate assay to detect MG in urine and anal samples.