Nanotube assisted microwave electroporation for single cell pathogen identification and antimicrobial susceptibility testing

Jian Gao; Hui Li; Peter Torab; Kathleen E Mach; David W Craft; Neal J Thomas; Chris M Puleo; Joseph C Liao; Tza-Huei Wang; Pak Kin Wong

doi:10.1016/j.nano.2019.01.015

Nanotube assisted microwave electroporation for single cell pathogen identification and antimicrobial susceptibility testing

Nanomedicine. 2019 Apr:17:246-253. doi: 10.1016/j.nano.2019.01.015. Epub 2019 Feb 20.

Authors

Jian Gao¹, Hui Li¹, Peter Torab², Kathleen E Mach³, David W Craft⁴, Neal J Thomas⁵, Chris M Puleo⁶, Joseph C Liao³, Tza-Huei Wang⁷, Pak Kin Wong⁸

Affiliations

¹ Department of Biomedical Engineering, The Pennsylvania State University, University Park, PA, USA.
² Department of Mechanical Engineering, The Pennsylvania State University, University Park, PA, USA.
³ Department of Urology, Stanford University School of Medicine, Stanford, CA, USA.
⁴ Departmemt of Pathology and Laboratory Medicine, Penn State Milton S. Hershey Medical Center, Hershey, PA, USA.
⁵ Departments of Pediatrics and Public Health Sciences, Penn State University College of Medicine, Hershey, PA, USA.
⁶ GE Global Research, Niskayuna, NY, USA.
⁷ Departments of Mechanical Engineering and Biomedical Engineering, Johns Hopkins University, Baltimore, MD, USA.
⁸ Department of Biomedical Engineering, The Pennsylvania State University, University Park, PA, USA; Department of Mechanical Engineering, The Pennsylvania State University, University Park, PA, USA; Department of Surgery, Penn State Milton S. Hershey Medical Center, Hershey, PA, USA. Electronic address: pak@engr.psu.edu.

Abstract

A nanotube assisted microwave electroporation (NAME) technique is demonstrated for delivering molecular biosensors into viable bacteria for multiplex single cell pathogen identification to advance rapid diagnostics in clinical microbiology. Due to the small volume of a bacterial cell (~femtoliter), the intracellular concentration of the target molecule is high, which results in a strong signal for single cell detection without amplification. The NAME procedure can be completed in as little as 30 minutes and can achieve over 90% transformation efficiency. We demonstrate the feasibility of NAME for identifying clinical isolates of bloodborne and uropathogenic pathogens and detecting bacterial pathogens directly from patient's samples. In conjunction with a microfluidic single cell trapping technique, NAME allows single cell pathogen identification and antimicrobial susceptibility testing concurrently. Using this approach, the time for microbiological analysis reduces from days to hours, which will have a significant impact on the clinical management of bacterial infections.

Keywords: Antibiotic resistance; Bacteria; Infection; Microfluidic; Urinary tract infection.

Publication types

Research Support, N.I.H., Extramural

MeSH terms

Anti-Bacterial Agents / pharmacology
Bacteria / drug effects
Bacteria / isolation & purification*
Bacterial Infections / diagnosis*
Bacterial Infections / drug therapy
Bacterial Infections / microbiology
Electroporation / instrumentation*
Equipment Design
Humans
Microbial Sensitivity Tests / instrumentation
Microfluidic Analytical Techniques / instrumentation
Microwaves
Nanotubes / chemistry*
Single-Cell Analysis / instrumentation*

Substances

Anti-Bacterial Agents

Grants and funding

R01 AI117032/AI/NIAID NIH HHS/United States