Miniaturized Antimicrobial Susceptibility Test by Combining Concentration Gradient Generation and Rapid Cell Culturing
Abstract
:1. Introduction
2. Results and Discussion
2.1. Microchip Design
2.2. Characterization of Generated Concentration Gradient
2.3. MIC Determination Using the Standard Broth Microdilution (SBM) Method
2.4. MIC Determination Using Microchip AST
3. Experimental Section
3.1. Bacterial Culture
3.2. MIC Determination by SBM Method
3.3. Microchip Fabrication
3.4. Fluorescence Intensity Measurements
3.5. Compression Injection
3.6. MIC Determination by Microchip AST
3.7. Image Processing by ImageJ Software
4. Conclusions
Supplementary Files
Supplementary File 1Acknowledgments
Author Contributions
Conflicts of Interest
References
- Jorgensen, J.H.; Ferraro, M.J. Antimicrobial susceptibility testing: A review of general principles and contemporary practices. Clin. Infect. Dis. 2009, 49, 1749–1755. [Google Scholar] [CrossRef] [PubMed]
- Clinical and Laboratory Standards Institute (CLSI). Clinical and Laboratory Standards Institute (CLSI) Website. Available online: http://clsi.org (accessed on 1 July 2014).
- Chen, C.H.; Lu, Y.; Sin, M.L.Y.; Mach, K.E.; Zhang, D.D.; Gau, V.; Liao, J.C.; Wong, P.K. Antimicrobial susceptibility testing using high surface-to-volume ratio microchannels. Anal. Chem. 2010, 82, 1012–1019. [Google Scholar] [CrossRef] [PubMed]
- Choi, J.; Jung, Y.-G.; Kim, J.; Kim, S.; Jung, Y.; Na, H.; Kwon, S. Rapid antibiotic susceptibility testing by tracking single cell growth in a microfluidic agarose channel system. Lab Chip 2012, 13, 280–287. [Google Scholar] [CrossRef] [PubMed]
- Hou, Z.; An, Y.; Hjort, K.; Hjort, K.; Sandegren, L.; Wu, Z. Time lapse investigation of antibiotic susceptibility using a microfluidic linear gradient 3D culture device. Lab Chip 2014, 14, 3409–3418. [Google Scholar] [CrossRef] [PubMed]
- Eun, Y.-J.; Utada, A.S.; Copeland, M.F.; Takeuchi, S.; Weibel, D.B. Encapsulating bacteria in agarose microparticles using microfluidics for high-throughput cell analysis and isolation. ACS Chem. Biol. 2011, 6, 260–266. [Google Scholar] [CrossRef] [PubMed]
- Boedicker, J.Q.; Li, L.; Kline, T.R.; Ismagilov, R.F. Detecting bacteria and determining their susceptibility to antibiotics by stochastic confinement in nanoliter droplets using plug-based microfluidics. Lab Chip 2008, 8, 1265–1272. [Google Scholar] [CrossRef] [PubMed]
- Mach, K.E.; Mohan, R.; Baron, E.J.; Shih, M.-C.; Gau, V.; Wong, P.K.; Liao, J.C. A biosensor platform for rapid antimicrobial susceptibility testing directly from clinical samples. J. Urol. 2011, 185, 148–153. [Google Scholar] [CrossRef] [PubMed]
- Tang, Y.; Zhen, L.; Liu, J.; Wu, J. Rapid antibiotic susceptibility testing in a microfluidic pH sensor. Anal. Chem. 2013, 85, 2787–2794. [Google Scholar] [CrossRef] [PubMed]
- Cira, N.J.; Ho, J.Y.; Dueck, M.E.; Weibel, D.B. A self-loading microfluidic device for determining the minimum inhibitory concentration of antibiotics. Lab Chip 2012, 12, 1052–1059. [Google Scholar] [CrossRef] [PubMed]
- Sinn, I.; Kinnunen, P.; Albertson, T.; McNaughton, B.H.; Newton, D.W.; Burns, M.A.; Kopelman, R. Asynchronous magnetic bead rotation (AMBR) biosensor in microfluidic droplets for rapid bacterial growth and susceptibility measurements. Lab. Chip 2011, 11, 2604–2611. [Google Scholar] [CrossRef] [PubMed]
- Mohan, R.; Mukherjee, A.; Sevgen, S.E.; Sanpitakseree, C.; Lee, J.; Schroeder, C.M.; Kenis, P.J.A. A multiplexed microfluidic platform for rapid antibiotic susceptibility testing. Biosens. Bioelectron. 2013, 49, 118–125. [Google Scholar] [CrossRef] [PubMed]
- Lu, Y.; Gao, J.; Zhang, D.D.; Gau, V.; Liao, J.C.; Wong, P.K. Single cell antimicrobial susceptibility testing by confined microchannels and electrokinetic loading. Anal. Chem. 2013, 85, 3971–3976. [Google Scholar] [CrossRef] [PubMed]
- Takagi, R.; Fukuda, J.; Nagata, K.; Yawata, Y.; Nomura, N.; Suzuki, H. A microfluidic microbial culture device for rapid determination of the minimum inhibitory concentration of antibiotics. Analyst 2013, 138, 1000–1003. [Google Scholar] [CrossRef] [PubMed]
- Price, C.S.; Kon, S.E.; Metzger, S. Rapid antibiotic susceptibility phenotypic characterization of Staphylococcus aureus using automated microscopy of small numbers of cells. J. Microbiol. Methods 2014, 98, 50–58. [Google Scholar] [CrossRef] [PubMed]
- Burnham, C.-A.D.; Frobel, R.A.; Herrera, M.L.; Wickes, B.L. Rapid ertapenem susceptibility testing and Klebsiella pneumoniae carbapenemase phenotype detection in Klebsiella pneumoniae isolates by use of automated microscopy of immobilized live bacterial cells. J. Clin. Microbiol. 2014, 52, 982–986. [Google Scholar] [CrossRef] [PubMed]
- Douglas, I.S.; Price, C.S.; Overdier, K.H.; Wolken, R.F.; Metzger, S.W.; Hance, K.R.; Howson, D.C. Rapid automated microscopy for microbiological surveillance of ventilator-associated pneumonia. Am. J. Respir. Crit. Care Med. 2015, 191, 566–573. [Google Scholar] [CrossRef] [PubMed]
- Choi, J.; Yoo, J.; Lee, M.; Kim, E.-G.; Lee, J.S.; Lee, S.; Joo, S.; Song, S.H.; Kim, E.-C.; Lee, J.C.; et al. A rapid antimicrobial susceptibility test based on single-cell morphological analysis. Sci. Transl. Med. 2014, 6. [Google Scholar] [CrossRef] [PubMed]
- Hattori, K.; Sugiura, S.; Kanamori, T. Generation of arbitrary monotonic concentration profiles by a serial dilution microfluidic network composed of microchannels with a high fluidic-resistance ratio. Lab Chip 2009, 9, 1763–1772. [Google Scholar] [CrossRef] [PubMed]
- Gervais, T.; El-Ali, J.; Günther, A.; Jensen, K.F. Flow-induced deformation of shallow microfluidic channels. Lab Chip 2006, 6, 500–507. [Google Scholar] [CrossRef] [PubMed]
- CLSI. Methods for Dilution Antimicrobial Susceptibility Tests for Bacteria that Grow Aerobically; Approved Standard—Seventh Edition (M7-A7); CLSI: Wayne, PA, USA, 2006; Volume 26, No. 2 CLSI M7A7. [Google Scholar]
- Kim, S.; Huang, B.; Zare, R.N. Microfluidic separation and capture of analytes for single-molecule spectroscopy. Lab Chip 2007, 7, 1663–1665. [Google Scholar] [CrossRef] [PubMed]
- Huang, B.; Wu, H.; Bhaya, D.; Grossman, A.; Granier, S.; Kobilka, B.K.; Zare, R.N. Counting low-copy number proteins in a single cell. Science 2007, 315, 81–84. [Google Scholar] [CrossRef] [PubMed]
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Kim, S.C.; Cestellos-Blanco, S.; Inoue, K.; Zare, R.N. Miniaturized Antimicrobial Susceptibility Test by Combining Concentration Gradient Generation and Rapid Cell Culturing. Antibiotics 2015, 4, 455-466. https://doi.org/10.3390/antibiotics4040455
Kim SC, Cestellos-Blanco S, Inoue K, Zare RN. Miniaturized Antimicrobial Susceptibility Test by Combining Concentration Gradient Generation and Rapid Cell Culturing. Antibiotics. 2015; 4(4):455-466. https://doi.org/10.3390/antibiotics4040455
Chicago/Turabian StyleKim, Samuel C., Stefano Cestellos-Blanco, Keisuke Inoue, and Richard N. Zare. 2015. "Miniaturized Antimicrobial Susceptibility Test by Combining Concentration Gradient Generation and Rapid Cell Culturing" Antibiotics 4, no. 4: 455-466. https://doi.org/10.3390/antibiotics4040455