Stepwise Evolution of Triple Antimicrobial Resistance in Neisseria meningitidis.

Abstract

Invasive meningococcal disease is a rapidly progressive infection with high mortality, for which effective antimicrobial therapy and chemoprophylaxis are essential. During national surveillance in El Salvador, invasive Neisseria meningitidis serogroup Y isolates with resistance to first-line agents were identified, prompting further investigation. Ten invasive N. meningitidis serogroup Y isolates, selected for dual resistance to penicillin and ciprofloxacin and collected between 2020 and 2025 by the National Public Health Laboratory of El Salvador, were analyzed at the Adolfo Lutz Institute, São Paulo, Brazil. Antimicrobial susceptibility testing was performed by minimum inhibitory concentration determination, and resistance mechanisms were characterized by whole-genome sequencing. All isolates were resistant to penicillin and ciprofloxacin. Genomic analysis showed that penicillin resistance was mediated by acquisition of the β-lactamase gene blaROB-1, while ciprofloxacin resistance was associated with a threonine-to-isoleucine substitution at position 91 of DNA gyrase (gyrA T91I). Two isolates additionally carried the tetB gene, defining a triple-resistance phenotype. These isolates exhibited markedly elevated tetracycline minimum inhibitory concentrations (6 μg/mL), approximately 40-fold higher than those observed in tetracycline-susceptible isolates. Increased MICs were also observed for minocycline and doxycycline, whereas susceptibility to newer tetracycline derivatives, including tigecycline and eravacycline, was preserved. All isolates belonged to sequence type 3587 within clonal complex 23. We report the first identification of N. meningitidis with concurrent resistance to penicillin, ciprofloxacin, and tetracycline, mediated by blaROB-1, gyrA T91I, and tetB, representing a critical escalation in meningococcal antimicrobial resistance.