One hundred forty-three penicillinase-producing Neisseria gonorrhoeae (PPNG) isolates obtained in Argentina from 2008 and 2012 were examined to detect bla TEM-135 genes and to investigate plasmid profiles and multiantigen sequence types. Forty-two PPNG isolates were found to carry TEM-135, and two contained a new TEM derivative characterized as TEM-220. The bla TEM-135 allele was carried by the Toronto/Rio and African plasmids. Molecular epidemiology revealed that two bla TEM-135 isolates were related to previously described isolates from Thailand and China, indicating a common evolutionary origin.A ntibiotic resistance in Neisseria gonorrhoeae is a global public health problem (1). Through the years, N. gonorrhoeae has developed resistance to the first-line antibiotics used for treatment (2). Penicillinase-producing N. gonorrhoeae (PPNG) isolates with plasmid-mediated high-level resistance to penicillin were first reported in 1976 and have spread since then (3). These first PPNG isolates contained TEM-1-type -lactamase plasmids encoded by transposon TnA (Tn2), which are responsible for the transference and dissemination of resistance (4). To date, eight plasmid types have been described in N. gonorrhoeae and named after their epidemiological origin as African, Toronto/Rio, Asian, Nîmes, New Zealand, Johannesburg, and Australian plasmids (5-7). The PPNG isolates carrying a bla TEM-135 gene were found in Thailand, Japan, and China and recently reported in Australia and 15 other countries (8-12). TEM-135 differs from TEM-1 by a single nucleotide substitution at position 539, resulting in a single amino acid substitution, M182T. This substitution is also found in TEM-type extended-spectrum -lactamase (ESBL) but has little effect on enzyme activity, and it has been proposed to stabilize substitutions near the active site, which collaboratively results in the emergence of a stable ESBL (13-15). The first PPNG isolate in Argentina was reported in 1980 and has since been disseminated in our country (16). The prevalence of PPNG isolates has been increasing through the years, with the highest level of 40% in the 1990's. The aim of this study was to detect bla TEM-135 and investigate plasmid types carrying -lactamase in the PPNG strains in 2008 and 2012 in Argentina.We studied 49 and 94 PPNG isolates collected in 2008 and 2012, respectively, from GASSP-AR, which includes 70 laboratories distributed all around the country. The MIC S (g/ml) of penicillin, ciprofloxacin, ceftriaxone, and azithromycin for the isolates were determined by the agar dilution method (17, 18). The N. gonorrhoeae ATCC 49226 and World Health Organization (WHO) reference strains were used for quality control in antimicrobial susceptibility testing (19). Mismatch amplification mutation assay (MAMA) PCR was performed to identify the bla TEM-135 allele, and TEM PCR was used to recognize both the bla TEM-1 and bla . The whole bla TEM , porB, and tbpB genes of all isolates that were MAMA PCR positive were amplified and sequenced to confirm the bla TEM-1...