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Research Article

Rapid phenotypic antimicrobial susceptibility testing using nanoliter arrays

Jonathan Avesar, Dekel Rosenfeld, Marianna Truman-Rosentsvit, Tom Ben-Arye, Yuval Geffen, Moran Bercovici, and Shulamit Levenberg
  1. aFaculty of Biomedical Engineering, Technion – Israel Institute of Technology, Haifa 3200003, Israel;
  2. bFaculty of Mechanical Engineering, Technion – Israel Institute of Technology, Haifa 3200003, Israel;
  3. cRussell Berrie Nanotechnology Institute, Technion – Israel Institute of Technology, Haifa 3200003, Israel;
  4. dMicrobiology Laboratory, Rambam Health Care Campus, Haifa 3109601, Israel

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PNAS first published June 26, 2017; https://doi.org/10.1073/pnas.1703736114
Jonathan Avesar
aFaculty of Biomedical Engineering, Technion – Israel Institute of Technology, Haifa 3200003, Israel;
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Dekel Rosenfeld
aFaculty of Biomedical Engineering, Technion – Israel Institute of Technology, Haifa 3200003, Israel;
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Marianna Truman-Rosentsvit
bFaculty of Mechanical Engineering, Technion – Israel Institute of Technology, Haifa 3200003, Israel;
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Tom Ben-Arye
aFaculty of Biomedical Engineering, Technion – Israel Institute of Technology, Haifa 3200003, Israel;
cRussell Berrie Nanotechnology Institute, Technion – Israel Institute of Technology, Haifa 3200003, Israel;
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Yuval Geffen
dMicrobiology Laboratory, Rambam Health Care Campus, Haifa 3109601, Israel
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Moran Bercovici
bFaculty of Mechanical Engineering, Technion – Israel Institute of Technology, Haifa 3200003, Israel;
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Shulamit Levenberg
aFaculty of Biomedical Engineering, Technion – Israel Institute of Technology, Haifa 3200003, Israel;
cRussell Berrie Nanotechnology Institute, Technion – Israel Institute of Technology, Haifa 3200003, Israel;
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  • For correspondence: shulamit@bm.technion.ac.il
  1. Edited by Robert Langer, Massachusetts Institute of Technology, Cambridge, MA, and approved June 1, 2017 (received for review March 6, 2017)

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Significance

Antibiotic resistance is fueled by antibiotic misuse and has become a major global health concern. The phenomenon warrants improved diagnostics that can more rapidly and efficiently elucidate information about the infectious agent to aid in establishing a more targeted and knowledge-based treatment regimen. This paper introduces a rapid antibiotic susceptibility test and automated data analysis algorithm that can, unlike traditional methods, deliver results on the same working day in an efficient and translatable manner for clinical use. This paper also introduces a method for direct urine testing that can help save days of diagnosis time. The platform is expected to promote more judicious use of antibiotics, thereby reducing the emergence of antibiotic resistance, lowering healthcare costs and ultimately saving lives.

Abstract

Antibiotic resistance is a major global health concern that requires action across all sectors of society. In particular, to allow conservative and effective use of antibiotics clinical settings require better diagnostic tools that provide rapid determination of antimicrobial susceptibility. We present a method for rapid and scalable antimicrobial susceptibility testing using stationary nanoliter droplet arrays that is capable of delivering results in approximately half the time of conventional methods, allowing its results to be used the same working day. In addition, we present an algorithm for automated data analysis and a multiplexing system promoting practicality and translatability for clinical settings. We test the efficacy of our approach on numerous clinical isolates and demonstrate a 2-d reduction in diagnostic time when testing bacteria isolated directly from urine samples.

  • antibiotic resistance
  • nanoliter wells
  • antibiotic susceptibility testing
  • microfluidics
  • resazurin

Footnotes

  • ↵1To whom correspondence should be addressed. Email: shulamit{at}bm.technion.ac.il.
  • Author contributions: J.A., D.R., T.B.A., Y.G., M.B., and S.L. designed research; J.A. and D.R. performed research; M.T.-R. contributed new reagents/analytic tools; J.A. analyzed data; J.A., D.R., T.B.A., Y.G., M.B., and S.L. wrote the paper; J.A. developed the analysis algorithm; T.B.A. helped design the analysis algorithm; and Y.G. guided the clinical work.

  • The authors declare no conflict of interest.

  • This article is a PNAS Direct Submission.

  • This article contains supporting information online at www.pnas.org/lookup/suppl/doi:10.1073/pnas.1703736114/-/DCSupplemental.

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Rapid phenotypic AST using nanoliter arrays
Jonathan Avesar, Dekel Rosenfeld, Marianna Truman-Rosentsvit, Tom Ben-Arye, Yuval Geffen, Moran Bercovici, Shulamit Levenberg
Proceedings of the National Academy of Sciences Jun 2017, 201703736; DOI: 10.1073/pnas.1703736114

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Rapid phenotypic AST using nanoliter arrays
Jonathan Avesar, Dekel Rosenfeld, Marianna Truman-Rosentsvit, Tom Ben-Arye, Yuval Geffen, Moran Bercovici, Shulamit Levenberg
Proceedings of the National Academy of Sciences Jun 2017, 201703736; DOI: 10.1073/pnas.1703736114
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Proceedings of the National Academy of Sciences: 118 (9)
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  • Article
    • Abstract
    • Description of the SNDA–AST System
    • Principle of Operation
    • Time to Results and Data Analysis
    • Antibiotic Incorporation via lyophilization and Multiplexing
    • Same-Day Detection and AST for UTIs
    • Discussion
    • Methods
    • Acknowledgments
    • Footnotes
    • References
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