Neisseria gonorrhoeae antimicrobial resistance (AMR) has been declared an urgent threat by the United States Centers for Disease Control and Prevention . Ceftriaxone is the mainstay of treatment, however many gonococcal strains throughout the world now exhibit reduced susceptibility, and in a small number of cases resistance, to this last recommended treatment. Emerging ceftriaxone-resistance in N. gonorrhoeae is primarily attributed to alterations in the penA gene which codes for penicillin binding protein 2 (PBP2). Of particular note is a N. gonorrhoeae strain harbouring a “mosaic” PBP2 sequence and which is of multi-locus sequence type 1901. This ‘mPBP2-1901’ strain now has a global distribution, typically exhibits reduced ceftriaxone susceptibility and is considered the precursor to the ceftriaxone-resistant F89 strain recently observed in Europe. In this study we sought to examine the prevalence of the ‘mPBP2-1901’ strain in Australia. The study was conducted as part of a NHMRC-funded study titled GRAND (Gonorrhoea Resistance Assessment via Nucleic acid Detection). A total of 2228 N. gonorrhoeae isolates collected in the first half of 2012, and estimated to comprise approximately 90% of all gonococci isolated in Australia in this time period, were characterized using the Sequenom iPLEX platform to provide MLST and AMR mutation data. Strikingly, we identified that the ‘mPBP2-1901’ strain comprised 8.4% (188/2228) of isolates in Australia in this time period. By State, the strain comprised 12%, 10.2%, 4.3% 1.5% and 0.6% of isolates in Victoria, New South Wales, Queensland, Western Australia and South Australia, respectively. Overall these data highlight the vulnerability of our population to importation and establishment of novel and potentially treatment-resistant gonococcal strains.