Limitations in predicting reduced susceptibility to third generation cephalosporins in Escherichia coli based on whole genome sequence data

PLoS One. 2023 Nov 30;18(11):e0295233. doi: 10.1371/journal.pone.0295233. eCollection 2023.

Abstract

Prediction of antibiotic resistance from whole genome sequence (WGS) data has been proposed. However, the performance of WGS data analysis for this matter may be influenced by the resistance mechanism's biology. This study compared traditional antimicrobial susceptibility testing with whole genome sequencing for identification of extended-spectrum beta-lactamases (ESBL) in a collection of 419 Escherichia coli isolates. BLASTn-based prediction and read mapping with srst2 gave matching results, and in 381/419 (91%) isolates WGS was congruent with phenotypic testing. Incongruent results were grouped by potential explanations into biological-related and sequence analysis-related results. Biological-related explanations included weak ESBL-enzyme activity (n = 4), inconclusive phenotypic ESBL-testing (n = 4), potential loss of plasmid during subculturing (n = 7), and other resistance mechanisms than ESBL-enzymes (n = 2). Sequence analysis-related explanations were cut-off dependency for read depth (n = 5), too stringent (n = 3) and too loose cut-off for nucleotide identity and coverage (n = 13), respectively. The results reveal limitations of both traditional antibiotic susceptibility testing and sequence-based resistance prediction and highlight the need for evidence-based standards in sequence analysis.

MeSH terms

  • Anti-Bacterial Agents / pharmacology
  • Anti-Bacterial Agents / therapeutic use
  • Cephalosporins / pharmacology
  • Escherichia coli Infections* / drug therapy
  • Escherichia coli*
  • Humans
  • Microbial Sensitivity Tests
  • beta-Lactamases / genetics

Substances

  • Anti-Bacterial Agents
  • beta-Lactamases
  • Cephalosporins

Grants and funding

SS holds a grant from ALF funds (ALF 810901) and the Swedish Research Council (2019-05909), and AH holds a grant from the Center for Research and Development Gävleborg (CFUG-698631). Generation of sequence data was supported by a grant from Afa Insurance, Sweden (grant number 150411) and Alf-de-Ruvo memorial foundation, Sweden. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.