Development and evaluation of a direct disk diffusion, rapid antimicrobial susceptibility testing method from blood culture positive for Gram-negative bacilli using rapid molecular testing and microbiology laboratory automation Journal Article
| Authors: | Cintrón, M.; Clark, B.; Miranda, E.; Delgado, M.; Esther Babady, N. |
| Article Title: | Development and evaluation of a direct disk diffusion, rapid antimicrobial susceptibility testing method from blood culture positive for Gram-negative bacilli using rapid molecular testing and microbiology laboratory automation |
| Abstract: | Rapid antimicrobial susceptibility testing (RAST) using disk diffusion (DD) from positive blood cultures (BC) can aid initiation of effective antimicrobial therapy. Rapid molecular blood culture identification panels (BCID) and microbiology laboratory automation (MLA) provide an opportunity to optimize RAST. This study aimed to evaluate and optimize RAST in combination with BCID and MLA for Escherichia coli, Klebsiella pneumoniae, and Pseudomonas aeruginosa positive BC. Monomicrobial BC positive for E. coli, K. pneumoniae, and P. aeruginosa by BCID between February 2020 and June 2021 were included. Images were captured by the MLA system after 4, 6, and 8 hours of incubation. The European Committee on Antimicrobial Susceptibility Testing (EUCAST) RAST breakpoints were used to interpret the zones of inhibitions measured by the WASPLab Halo Recognition software. Categorical agreement (CA) and error rates (minor [mE], major [ME] and very major [VME] errors) were calculated using automated microbroth dilution (aMBD) and DD as comparator methods. Ninety-one BCs met the inclusion criteria. CA between RAST and aMBD was >90% for most antimicrobials tested at all three time points for E. coli and K. pneumoniae. Overall, mE rates were <4%, ME rates ranged from 1% to 6% and VME rates ranged from 2% to 9%. Performance of RAST was lowest for piperacillin/tazobactam (71%–86%). For P. aeruginosa, CAs were mostly <90% with significant error rates. Combining rapid organism identification by BCID with automated RAST set-up and interpretation can facilitate the generation of early, preliminary antimicrobial susceptibility results for E. coli and K. pneumoniae. Copyright © 2025 Cintrón et al. |
| Keywords: | genetics; drug effect; antiinfective agent; anti-bacterial agents; artificial intelligence; e. coli; escherichia coli; microbiology; isolation and purification; gram-negative bacteria; gram-negative bacterial infections; pseudomonas aeruginosa; blood culture; klebsiella pneumoniae; procedures; gram negative infection; gram negative bacterium; microbial sensitivity tests; laboratory automation; automation, laboratory; disk diffusion; humans; human; evaluation study; microbial sensitivity test; eucast; klebsiella pneumoniae and pseudomonas aeruginosa; rapid antimicrobial susceptibility testing; disk diffusion antimicrobial tests |
| Journal Title: | Microbiology Spectrum |
| Volume: | 13 |
| Issue: | 6 |
| ISSN: | 2165-0497 |
| Publisher: | American Society for Microbiology |
| Date Published: | 2025-06-01 |
| Start Page: | e0240124 |
| Language: | English |
| DOI: | 10.1128/spectrum.02401-24 |
| PUBMED: | 40372037 |
| PROVIDER: | scopus |
| PMCID: | PMC12131842 |
| DOI/URL: | |
| Notes: | Article -- MSK Cancer Center Support Grant (P30 CA008738) acknowledged in PubMed and PDF -- MSK corresponding author is N. Esther Babady -- Source: Scopus |
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