Antibiotic susceptibility testing in less than 30 min using direct single-cell imaging

Proc Natl Acad Sci U S A. 2017 Aug 22;114(34):9170-9175. doi: 10.1073/pnas.1708558114. Epub 2017 Aug 8.

Abstract

The emergence and spread of antibiotic-resistant bacteria are aggravated by incorrect prescription and use of antibiotics. A core problem is that there is no sufficiently fast diagnostic test to guide correct antibiotic prescription at the point of care. Here, we investigate if it is possible to develop a point-of-care susceptibility test for urinary tract infection, a disease that 100 million women suffer from annually and that exhibits widespread antibiotic resistance. We capture bacterial cells directly from samples with low bacterial counts (104 cfu/mL) using a custom-designed microfluidic chip and monitor their individual growth rates using microscopy. By averaging the growth rate response to an antibiotic over many individual cells, we can push the detection time to the biological response time of the bacteria. We find that it is possible to detect changes in growth rate in response to each of nine antibiotics that are used to treat urinary tract infections in minutes. In a test of 49 clinical uropathogenic Escherichia coli (UPEC) isolates, all were correctly classified as susceptible or resistant to ciprofloxacin in less than 10 min. The total time for antibiotic susceptibility testing, from loading of sample to diagnostic readout, is less than 30 min, which allows the development of a point-of-care test that can guide correct treatment of urinary tract infection.

Keywords: AST; UTI; antibiotic; microfluidic; point of care; resistance.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Anti-Bacterial Agents / pharmacology*
  • Ciprofloxacin / pharmacology
  • Drug Resistance, Microbial / drug effects
  • Escherichia coli Infections / microbiology
  • Female
  • Humans
  • Microbial Sensitivity Tests / methods*
  • Point-of-Care Testing / standards
  • Reproducibility of Results
  • Single-Cell Analysis / methods*
  • Urinary Tract Infections / microbiology
  • Uropathogenic Escherichia coli / classification
  • Uropathogenic Escherichia coli / drug effects*
  • Uropathogenic Escherichia coli / physiology

Substances

  • Anti-Bacterial Agents
  • Ciprofloxacin