With the aim of studying the dynamics of TPA strains in Buenos Aires, Argentina, the purpose of the present study was to characterize the syphilis epidemic in Buenos Aires by analyzing TPA clinical strains collected during 2015–2019 in terms of their strain and haplotype distribution as well as the presence of antibiotic resistance-associated mutations, in order to update and expand on our previous report using the newer MLST methodology. The gene discussed is PLAT; the disease is syphilis.