Single cell antimicrobial susceptibility testing by confined microchannels and electrokinetic loading

Yi Lu, Jian Gao, Donna D. Zhang, Vincent Gau, Joseph C. Liao, Pak Kin Wong

Research output: Contribution to journalArticle

59 Scopus citations

Abstract

Multidrug-resistant pathogens are an emerging global health problem. In addition to the need of developing new antibiotics in the pipeline, the ability to rapidly determine the antibiotic resistance profiles of bacteria represents one of the most crucial steps toward the management of infectious diseases and the prevention of multidrug-resistant pathogens. Here, we report a single cell antimicrobial susceptibility testing (AST) approach for rapid determination of the antibiotic resistance of bacterial pathogens. By confining individual bacteria in gas permeable microchannels with dimensions comparable to a single bacterium, the antibiotic resistance of the bacteria can be monitored in real-time at the single cell level. To facilitate the dynamic loading of the bacteria into the confined microchannels for observation, AC electrokinetics is demonstrated for capturing bacteria to defined locations in high-conductivity AST buffer. The electrokinetic technique achieves a loading efficiency of about 75% with a negligible effect on the bacterial growth rate. To optimize the protocol for single cell AST, the bacterial growth rate of individual bacteria under different antibiotic conditions has been determined systematically. The applicability of single cell AST is demonstrated by the rapid determination of the antimicrobial resistant profiles of uropathogenic clinical isolates in Mueller-Hinton media and in urine. The antibiotic resistance profiles of bacteria can be determined in less than 1 h compared to days in standard culture-based AST techniques.

Original languageEnglish (US)
Pages (from-to)3971-3976
Number of pages6
JournalAnalytical chemistry
Volume85
Issue number8
DOIs
StatePublished - Apr 16 2013

ASJC Scopus subject areas

  • Analytical Chemistry

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