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Fluorescence Assay Based on Aptamer-Quantum Dot Binding to Bacillus thuringiensis Spores

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Abstract

A novel assay was developed for the detection of Bacillus thuringiensis (BT) spores. The assay is based on the fluorescence observed after binding an aptamer-quantum dot conjugate to BT spores. The in vitro selection and amplification technique called SELEX (Systematic Evolution of Ligands by EXponential enrichment) was used in order to identify the DNA aptamer sequence specific for BT. The 60 base aptamer was then coupled to fluorescent zinc sulfide-capped, cadmium selenide quantum dots (QD). The assay is semi-quantitative, specific and can detect BT at concentrations of about 1,000 colony forming units/ml.

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Acknowledgements

The authors thank the U.S. Air Force Research Laboratory, Brooks City-Base, San Antonio, TX, for partial laboratory support. We would also like to acknowledge the Department of Defense (U.S. Army Contract No. DACA42-03-C-0063) for financial support of this project.

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Author notes

  1. Amity Allman

    Present address: Severn Trent Laboratories, 880 Riverside Pwky, West Sacramento, CA, 95605, USA

Authors and Affiliations

  1. Department of Chemistry and Biochemistry, Texas State University-San Marcos, 601 University Drive, San Marcos, TX, 78666, USA

    Milada Ikanovic & Walter E. Rudzinski

  2. Operational Technologies Corporation, 4100 NW Loop 410, Suite 230, San Antonio, TX, 78229, USA

    John G. Bruno, Amity Allman & Maria P. Carrillo

  3. NanoScience Diagnostics, Inc., 1826 Kramer Lane, Suite E, Austin, TX, 78758, USA

    Sulatha Dwarakanath, Suneetha Bhahdigadi & Poornima Rao

  4. Air Force Research Laboratory, HEPC, 2486 Gillingham Drive, Bldg. 175 E, Brooks City-Base, San Antonio, TX, 78235, USA

    Johnathan L. Kiel & Carrie J. Andrews

Authors
  1. Milada Ikanovic
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  2. Walter E. Rudzinski
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  3. John G. Bruno
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  4. Amity Allman
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  5. Maria P. Carrillo
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  6. Sulatha Dwarakanath
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  7. Suneetha Bhahdigadi
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  8. Poornima Rao
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  9. Johnathan L. Kiel
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Correspondence to Walter E. Rudzinski.

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Ikanovic, M., Rudzinski, W.E., Bruno, J.G. et al. Fluorescence Assay Based on Aptamer-Quantum Dot Binding to Bacillus thuringiensis Spores. J Fluoresc 17, 193–199 (2007). https://doi.org/10.1007/s10895-007-0158-4

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  • DOI: https://doi.org/10.1007/s10895-007-0158-4

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