Introduction: Mycoplasma genitalium (MG) is a sexually transmitted microbe associated with urethritis in men and several inflammatory syndromes in women. Macrolides and fluoroquinolones have been recommended treatments for MG infections. However, resistance of both drug classes is increasing worldwide and treatment failures have been described. There are very little data available on antimicrobial resistance in MG from the USA.

Methods We investigated the prevalence of macrolide- and fluoroquinolone-resistant MG in African American men and women who presented to a Sexually Transmitted Diseases Clinic in Birmingham, Alabama during 2015–2016; most were couples. A real-time PCR assay was validated for detecting MG 23S rRNA mutations known to confer macrolide resistance directly from clinical specimens. Two nested PCRs were used to detect mutations in the quinolone resistant determination regions (QRDRs) in gyrA and parC genes.

Results Oral, rectal, urine, and/or vaginal specimens from 90 men and 81 women have been tested thus far. A total of 23 MG-positive patients have been identified (4 couples and 15 singles), giving a prevalence rate for MG of 13.5% in this cohort; 11 (12.2%) of men and 12 (14.8%) of women were MG positive. Eleven (47.8%) patients (6 men and 5 women) carried macrolide-resistant MG. Sequencing of the PCR products indicated that A2071G and A2072G transitions in the 23S rRNA gene was the major mutations. No mutation was found in gyrA QRDR. Three patients (13.6%), including 1 couple, carried G248T (S83I) mutation in parC QRDR. The female of the couple also carried a parC G259C (D87H) mutant group. Two individuals (a couple) carried organisms that had markers for both macrolide and fluoroquinolone resistance.

Conclusion This is the first study based in the USA to document both fluoroquinolone and macrolide resistance in MG using a molecular-based assay. We plan to genotype MG directly from the clinical specimens to investigate the genetic relatedness of antimicrobial-susceptible and resistant-MG, as well as the concordance of MG in couples.