Background:
Neisseria gonorrhoeae has been identified by the CDC as an urgent antimicrobial threat. Similarly, the World Health Organisation has identified N. gonorrhoeae as a priority for enhanced, quality-assured antimicrobial resistance (AMR) surveillance. Current point-of-care diagnostics for Gonorrhea are not widely available and currently cannot inform antimicrobial selection. Therefore, detection of resistance and decreased susceptibility in circulating strains is critical to maintain effective antimicrobial stewardship. To fulfill this aim, a molecular AMR screen for N. gonorrhoeae was developed.
Methods:
In collaboration with New Zealand and Queensland government health departments, primers targetting 15 genes from NG-MAST, Neisseria MLST, and NG-STAR typing schemes were utilised, alongside primers targetting plasmid-mediated penicillin resistance. Amplicon-based gene targets were selected to improve cost effectiveness compared to whole genome sequencing, and were amplified in a multiplexed manner for efficiency. Validation in silico and through wet lab verification against local and global N. gonorrhoeae sequences is ongoing.
Results:
An automated sequencing pipeline was established to process NextSeq Illumina sequencing and implement downstream bioinformatic analysis, specifically looking for N. gonorrhoeae antimicrobial resistance markers. Submission into the globally maintained typing schemes allows prediction of resistance phenotype, and identification of novel resistance types.
Conclusion:
This work demonstrates viability and utility of a broad but affordable sexually transmitted infection AMR surveillance pipeline. This will be implemented into routine use as part of Queensland Health’s molecular surveillance of N. gonorrhoeae isolates. This pipeline will inform on emerging N. gonorrhoeae resistance to major antibiotics in Queensland, and underpin enhanced antimicrobial stewardship to preserve antibiotics.