Molecular characterization of Neisseria gonorrhoeae isolates collected through a national surveillance programme in Japan, 2013: evidence of the emergence of a ceftriaxone-resistant strain from a ceftriaxone-susceptible lineage

Abstract

Objectives To investigate the spread of ceftriaxone-resistant Neisseria gonorrhoeae lineages similar to strains H041 (2009) and FC428 (2015), we characterized 55 strains collected in 2013 from hospitals across Japan. Methods Susceptibility testing and whole-genome sequencing. Results Susceptibility rates were 58% for cefixime and 98% for ceftriaxone. The 55 strains were whole-genome sequenced and classified into nine MLST-STs. MLST-ST1901 was the most prevalent (n = 19) followed by MLST-ST7363 (n = 12) and MLST-ST7359 (n = 11). The most prevalent penA [encoding penicillin binding protein 2 (PBP2)] mosaic types, based on the N. gonorrhoeae sequence typing for antimicrobial resistance (NG-STAR) scheme, were 10.001 (n = 20) followed by 34.001 (n = 13). The H041 and FC428 strains were not detected; however, a single ceftriaxone-resistant strain (TUM15748) with a MIC of 0.5 mg/L ceftriaxone was identified. The TUM15748 strain belonged to MLST-ST7359 and N. gonorrhoeae multiantigen sequence typing-ST6771, and had a novel PBP2 (PBP2TUM15748, penA type 169.001). The amino acid sequence of PBP2TUM15748 showed partial similarity to that of PBP2 from N. gonorrhoeae GU140106 and commensal Neisseria perflava and Neisseria cinerea. Natural transformation and recombination experiments using full-length TUM15748 penA showed that the ceftriaxone MICs of transformants increased 16-fold or more compared with the parental ceftriaxone-susceptible recipient strain (NG9807, belonging to MLST-ST7363). No ceftriaxone-resistant MLST-ST7359 strains have previously been reported. Conclusions We showed here that a ceftriaxone-susceptible lineage acquired a mutant PBP2 mosaic type, integrating partial PBP2 sequences from commensal Neisseria species, resulting in the emergence of ceftriaxone-resistant strains.

Genetic Environment Surrounding bla OXA-55-like in Clinical Isolates of Shewanella algae Clade and Enhanced Expression of bla OXA-55-like in a Carbapenem-Resistant Isolate

“…Additionally, long-read nucleotide sequences were obtained using the MinION sequencer (Oxford Nanopore Technologies, Oxford, United Kingdom) for isolates identified as S. algae clade members ( S. algae , S. carassii , or S. chilikensis ) to determine the complete whole-genome sequences. DNA extraction, library preparation, sequencing, and de novo assembly for MinION were performed as previously published ( 17 ).…”

Development and validation of a high throughputNeisseria gonorrhoeaegenotyping method

“…Because Japan has been an epicenter for AMR N. gonorrhoeae 12,15,[31][32][33][34] , it is appropriate to use Japanese strains to validate our typing methodology.…”

“…Whole-genome sequencing (WGS) using a massive parallel sequencer (MPS) can simultaneously analyze multiple typing methods 15 , such as multilocus sequence typing (MLST) 16,17 , N. gonorrhoeae multi-antigen sequence typing (NG-MAST) 18 , and N. gonorrhoeae sequence typing for antimicrobial resistance (NG-STAR) scheme for 7 alleles associated with cephalosporins, fluoroquinolones, penicillins, and macrolides 19 . WGS using MPS is widely used and allows for a comprehensive analysis, but its cost per strain is too expensive for large-scale surveillance 15,18,20,21 . Hence, a less expensive and more efficient AMR N. gonorrhoeae genotyping method is required.…”

First National Genomic Epidemiological Study of Neisseria gonorrhoeae Strains Spreading Across Sweden in 2016

“…However, these STs have been less frequently observed in other EU/EEA countries. Limited observations have been made in the United Kingdom and United States (ST8156) and Japan (ST7359) but the majority of isolates has been found in Australia (ST8156 and ST7359) (Buckley et al, 2018;Hanao et al, 2021;Pathogenwatch, 2021). In 2013, MLST ST7363, ST1901 and ST1588 were the most prevalent STs in isolates from 20 different EU/EEA countries (Harris et al, 2018).…”

“…Cefiximeresistant MLST ST7363 strains frequently harbor mosaic penA-10. Both mosaic penA-10 and mosaic penA-34 have been described in ESC-resistant N. gonorrhoeae strains worldwide, e.g., in Japan, Vietnam, Argentina, United States, Canada and Europe (Demczuk et al, 2015;Grad et al, 2016;Harris et al, 2018;Gianecini et al, 2019;Lan et al, 2020;Hanao et al, 2021;Pathogenwatch, 2021). In Sweden, 85% of all isolates with mosaic penA alleles belonged to MLST ST1901 and ST7363 and these isolates were mostly associated with domestic heterosexual transmission.…”