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A liposome-PCR assay for the ultrasensitive detection of biological toxins

Nature Biotechnology volume 24, pages 555–557 (2006)Cite this article

Abstract

We describe an ultrasensitive immunoassay for detecting biotoxins that uses liposomes with encapsulated DNA reporters, and ganglioside receptors embedded in the bilayer, as a detection reagent. After immobilization of the target biotoxin by a capture antibody and co-binding of the detection reagent, the liposomes are ruptured to release the reporters, which are quantified by real-time PCR. Assays for cholera and botulinum toxins are several orders of magnitude more sensitive than current detection methods.

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Figure 1: Representation of a liposome detection reagent in cross section.
Figure 2: Plots of the average serial dilution Ct values versus the log of the number of molecules per plate well for the four replicate measurements of (a) CTBS in deionized water and (b) BoNT/A in deionized water.

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Acknowledgements

The authors thank Marilyn Mason for scientific advice and editorial assistance. This work was supported by Army Medical Research and Material Command Grant DAMD17-02-1-0178 to J.T.M. The opinions or assertions herein are those of the authors and do not necessarily reflect the view of the Department of the Army or the Department of Defense.

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Author notes

  1. Lixin Xu

    Present address: Division of Monoclonal Antibodies, Center for Drug Evaluation and Research, US Food and Drug Administration, 29 Lincoln Drive, Bethesda, Maryland, 20892, USA

  2. Lixin Xu and Zong-mei Sheng: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Biophysics, Armed Forces Institute of Pathology, 1413 Research Boulevard, Rockville, 20850, Maryland, USA

    Jeffrey T Mason & Lixin Xu

  2. Department of Molecular Pathology, Armed Forces Institute of Pathology, 1413 Research Boulevard, Rockville, 20850, Maryland, USA

    Zong-mei Sheng

  3. Biomedical Laboratory Research and Development Service, Veterans Health Administration, 810 Vermont Avenue, NW, Washington, 20420, DC, USA

    Timothy J O'Leary

Authors
  1. Jeffrey T Mason
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  2. Lixin Xu
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  3. Zong-mei Sheng
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  4. Timothy J O'Leary
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Corresponding author

Correspondence to Jeffrey T Mason.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Fig. 1

Representative real-time PCR amplification curves. (PDF 23 kb)

Supplementary Fig. 2

LPCR assay of CTBS in run-off water. (PDF 65 kb)

Supplementary Fig. 3

LPCR assay of CTBS in human urine. (PDF 66 kb)

Supplementary Table 1

Comparison of biotoxin assay methods. (PDF 9 kb)

Supplementary Methods (PDF 118 kb)

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Mason, J., Xu, L., Sheng, Zm. et al. A liposome-PCR assay for the ultrasensitive detection of biological toxins. Nat Biotechnol 24, 555–557 (2006). https://doi.org/10.1038/nbt1201

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