Using a real-time PCR assay specific for a mosaic penA allele that has been associated with oral cephalosporin resistance in Asia, 54 available Neisseria gonorrhoeae isolates collected in San Francisco, CA, from January to October 2008 were analyzed. Five isolates tested positive for the mosaic penA gene by real-time PCR. DNA sequencing revealed two mosaic penA alleles (SF-A and SF-B). Isolates with SF-A and SF-B alleles possessed elevated MICs for the oral cephalosporins cefpodoxime and cefixime.

eThe recent emergence of Neisseria gonorrhoeae strains with decreased susceptibility to extended-spectrum cephalosporins is a major concern globally. We sequenced the genome of an N. gonorrhoeae multiantigen sequence typing (NG-MAST) ST1407 isolate (SM-3) with decreased susceptibility and resistance to oral extended-spectrum cephalosporins. The isolate was cultured in 2008 in San Francisco, CA, and possessed mosaic penA allele XXXIV, which is associated with an international clone that possesses decreased susceptibility as well as resistance to oral extended-spectrum cephalosporins globally. The genome sequence of strain NCCP11945 was used as a scaffold, and our assembly resulted in 91 contigs covering 2,029,064 bp (91%; >150؋ coverage) of the genome. Numerous instances of suspected horizontal genetic transfer events with other Neisseria species were identified, and two genes, opa and txf, acquired from nongonococcal Neisseria species, were identified. Strains possessing mosaic penA alleles (n ‫؍‬ 108) and nonmosaic penA alleles (n ‫؍‬ 169) from the United States and Europe (15 countries), cultured in 2002 to 2009, were screened for the presence of these genes. The opa gene was detected in most (82%) penA mosaic-containing isolates (mainly from 2007 to 2009) but not in any penA nonmosaic isolates. The txf gene was found in all strains containing opa but also in several (18%) penA nonmosaic strains. Using opa and txf as genetic markers, we identified a strain that possesses mosaic penA allele XXXIV, but the majority of its genome is not genetically related to strain SM-3. This implies that penA mosaic allele XXXIV was transferred horizontally. Such isolates also possessed decreased susceptibility and resistance to oral extended-spectrum cephalosporins. These findings support that genetic screening for particular penA mosaic alleles can be a valuable method for tracking strains with decreased susceptibility as well as resistance to oral extended-spectrum cephalosporins worldwide and that screening using only NG-MAST may not be sufficient.

Urogenital Neisseria gonorrhoeae isolates (266) collected in San Francisco, CA, in 2009 were analyzed for antimicrobial susceptibility and were subsequently genotyped by N. gonorrhoeae multiantigen sequence typing (NG-MAST). Isolates of identical or closely related sequence types were found to possess highly similar phenotypes with regard to drug susceptibility. Isolates containing decreased susceptibility to oral cephalosporins were detected in 2009 and were found to contain the mosaic penA allele (XXXIV) found previously to be associated with decreased susceptibility to cephalosporins. A better understanding of the relationships between phenotypic and genotypic markers for antimicrobial resistance may be helpful to the development of effective surveillance systems for drug-resistant N. gonorrhoeae.

We identified strains of Neisseria gonorrhoeae in San Diego County, California, that possess high levels of resistance to azithromycin (8.0-16.0 μg/ml MIC). These isolates contain a combination of mutations in both the 23S rRNA loci and mtrR genes. This is the first description of such isolates in the United States.