Abstract
AbstractBackgroundNeisseria gonorrhoeaegenotyping by whole-genome sequencing (WGS) is expensive for a large sample set, a less expensive and more efficient genotyping method is required. We developed a high-throughput genotyping method forN. gonorrhoeaeto improve molecular epidemiological typing and antimicrobial-resistant identification inN. gonorrhoeaeantimicrobial susceptibility surveillance.MethodsWe used multiplex-tailed PCR to amplify and sequence 15 alleles from multilocus sequence typing (MLST),N. gonorrhoeaemultiantigen sequence typing (NG-MAST), andN. gonorrhoeaesequence typing for antimicrobial resistance (NG-STAR). After indexing-PCR, we sequenced the DNA library using the MiSeq platform (Illumina). Sequencing reads werede novoassembly or constructing consensus sequences of alleles, then assigned sequence type. We used 54 previously characterized strains ofN. gonorrhoeaeand WGS data to validate our method.ResultsThe allele identification results of MLST and NG-STAR in all strains agreed with the draft WGS. However, in NG-MAST, only 35 strains agreed. Disagreement was found in the NG-MAST ofporBin 15 strains and oftbpBin seven strains. QRDR analysis perfectly predicted levofloxacin resistance. But was less successful in predicting reduced susceptibility or resistance phenotype to penicillin G, cefixime, or ceftriaxone usingpenA, porB, ponA, ormtrRalleles.ConclusionsThe successful performance in MLST and NG-STAR of our method was validated in this study. This method may be useful for large-scale genotyping forN. gonorrhoeaesurveillance in a cost- and labor-saving manner. Phenotypic prediction of antimicrobial susceptibility by combining multiple alleles may be necessary for other than fluoroquinolones.
Publisher
Cold Spring Harbor Laboratory
Reference34 articles.
1. World Health Organization (WHO). Report on global sexually transmitted infection surveillance, 2018. 2018. Available at: https://www.who.int/publications/i/item/9789241565691 [verified January 2023].
2. Surveillance and epidemiology of syphilis, gonorrhoea and chlamydia in the non-European Union countries of the World Health Organization European Region, 2015 to 2020;Euro Surveill,2022
3. World Health Organization Global Gonococcal Antimicrobial Surveillance Program (WHO GASP): review of new data and evidence to inform international collaborative actions and research efforts;Sex Health,2019
4. Antimicrobial Resistance in Neisseria gonorrhoeae in the 21st Century: Past, Evolution, and Future
5. Optimising treatments for sexually transmitted infections: surveillance, pharmacokinetics and pharmacodynamics, therapeutic strategies, and molecular resistance prediction;Lancet Infect Dis,2020
Cited by
1 articles.
订阅此论文施引文献
订阅此论文施引文献,注册后可以免费订阅5篇论文的施引文献,订阅后可以查看论文全部施引文献