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Integration of spore-based genetically engineered whole-cell sensing systems into portable centrifugal microfluidic platforms

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Abstract

Bacterial whole-cell biosensing systems provide important information about the bioavailable amount of target analytes. They are characterized by high sensitivity and specificity/selectivity along with rapid response times and amenability to miniaturization as well as high-throughput analysis. Accordingly, they have been employed in various environmental and clinical applications. The use of spore-based sensing systems offers the unique advantage of long-term preservation of the sensing cells by taking advantage of the environmental resistance and ruggedness of bacterial spores. In this work, we have incorporated spore-based whole-cell sensing systems into centrifugal compact disk (CD) microfluidic platforms in order to develop a portable sensing system, which should enable the use of these hardy sensors for fast on-field analysis of compounds of interest. For that, we have employed two spore-based sensing systems for the detection of arsenite and zinc, respectively, and evaluated their analytical performance in the miniaturized microfluidic format. Furthermore, we have tested environmental and clinical samples on the CD microfluidic platforms using the spore-based sensors. Germination of spores and quantitative response to the analyte could be obtained in 2.5–3 h, depending on the sensing system, with detection limits of 1 × 10−7 M for arsenite and 1 × 10−6 M for zinc in both serum and fresh water samples. Incorporation of spore-based whole-cell biosensing systems on microfluidic platforms enabled the rapid and sensitive detection of the analytes and is expected to facilitate the on-site use of such sensing systems.

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Acknowledgments

This work was supported by the Superfund Research Program (SRP) of the National Institute of Environmental Health Sciences (NIEHS; grant P42ES07380), the National Science Foundation (NSF; grant CHE-0718844), and the United States–Israel Binational Agricultural Research and Development (BARD) Fund (grant US-3864-06). We would like to thank Dr. Tsutomu Sato (Tokyo University of Agriculture and Technology, Japan) for kindly providing the plasmid pMUTin-23 and the bacterial strain B. subtilis ars-23.

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  1. Department of Chemistry, University of Kentucky, Lexington, KY, 40506-0055, USA

    Amol Date, Patrizia Pasini & Sylvia Daunert

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  1. Amol Date
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  2. Patrizia Pasini
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  3. Sylvia Daunert
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Correspondence to Sylvia Daunert.

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Date, A., Pasini, P. & Daunert, S. Integration of spore-based genetically engineered whole-cell sensing systems into portable centrifugal microfluidic platforms. Anal Bioanal Chem 398, 349–356 (2010). https://doi.org/10.1007/s00216-010-3930-2

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  • DOI: https://doi.org/10.1007/s00216-010-3930-2

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