Bacterial nanoscale cultures for phenotypic multiplexed antibiotic susceptibility testing

Area of Science:

• Microbiology
• Medical Diagnostics
• Biotechnology

Background:

• Effective treatment of bacterial infections relies on optimal antimicrobial drug regimens.
• Rapid and precise antibiotic susceptibility profiling is crucial for guiding antibiotic selection.
• Current methods can be time-consuming, delaying critical treatment decisions.

Purpose of the Study:

• To develop a high-throughput nanowell device for rapid antibiotic susceptibility testing (AST).
• To enable direct, multiplexed analysis of bacterial antibiotic resistance.
• To reduce the time required for determining minimum inhibitory concentrations (MICs).

Main Methods:

• Utilized a high-throughput nanowell device for culturing bacterial strains.
• Employed real-time optical recordings of nanoscale cultures.
• Applied a mathematical algorithm to identify the growth shift from lag to early logarithmic phase (Tlag).
• Analyzed reference and clinical uropathogenic Escherichia coli strains.

Main Results:

• Successfully identified Tlag for hundreds of nanoscale cultures in real time.
• Defined the minimum inhibitory concentration (MIC) within 4 hours.
• Enabled rapid differentiation of multiple antibiotic resistance patterns using heatmap visualization.
• Demonstrated the device's capability for high-throughput diagnostic analysis.

Conclusions:

• The developed nanowell AST device provides a rapid and high-throughput method for antibiotic susceptibility profiling.
• This technology significantly reduces the time to result for MIC determination.
• The device aids clinicians in making timely and informed decisions for optimal antibiotic therapy selection.