Single cell antimicrobial susceptibility testing by confined microchannels and electrokinetic loading

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

Research output: Contribution to journalArticle

49 Citations (Scopus)

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

Fingerprint

Microchannels
Bacteria
Testing
Anti-Bacterial Agents
Pathogens
Medical problems
Buffers
Pipelines
Gases

ASJC Scopus subject areas

  • Analytical Chemistry

Cite this

Single cell antimicrobial susceptibility testing by confined microchannels and electrokinetic loading. / Lu, Yi; Gao, Jian; Zhang, Donna; Gau, Vincent; Liao, Joseph C.; Wong, Pak Kin.

In: Analytical Chemistry, Vol. 85, No. 8, 16.04.2013, p. 3971-3976.

Research output: Contribution to journalArticle

@article{e97b0c32ada14195a607c524be909562,
title = "Single cell antimicrobial susceptibility testing by confined microchannels and electrokinetic loading",
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.",
author = "Yi Lu and Jian Gao and Donna Zhang and Vincent Gau and Liao, {Joseph C.} and Wong, {Pak Kin}",
year = "2013",
month = "4",
day = "16",
doi = "10.1021/ac4004248",
language = "English (US)",
volume = "85",
pages = "3971--3976",
journal = "Analytical Chemistry",
issn = "0003-2700",
publisher = "American Chemical Society",
number = "8",

}

TY - JOUR

T1 - Single cell antimicrobial susceptibility testing by confined microchannels and electrokinetic loading

AU - Lu, Yi

AU - Gao, Jian

AU - Zhang, Donna

AU - Gau, Vincent

AU - Liao, Joseph C.

AU - Wong, Pak Kin

PY - 2013/4/16

Y1 - 2013/4/16

N2 - 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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=84876234983&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84876234983&partnerID=8YFLogxK

U2 - 10.1021/ac4004248

DO - 10.1021/ac4004248

M3 - Article

C2 - 23445209

AN - SCOPUS:84876234983

VL - 85

SP - 3971

EP - 3976

JO - Analytical Chemistry

JF - Analytical Chemistry

SN - 0003-2700

IS - 8

ER -