Micro-NMR and Nanoparticle Amplification                              for Botulinum Toxin                                              Diagnostics

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Genetic Screens to Identify the Ebola Virus Receptor

High-throughput Assay Development Against                              Cryptosporidium Glycotlytic                              Enzymes

Model for Oral Ingestion of Ricin Toxin

Micro-NMR and Nanoparticle Amplification for Botulinum Toxin Diagnostics

Collaborating Institution: Sandia National Laboratories, Albuquerque, NM

Principal Investigator: Todd M. Alam, Ph.D.

Co-Investigators:
a) David P. Adams, Ph.D. – Sandia National Laboratories, Albuquerque, NM
b) Laurel O. Sillerud, Ph.D. – University of New Mexico, Albuquerque, NM
c) John D. Williams, Ph.D. – Sandia National Laboratories, Albuquerque, NM

Expected Product: Diagnostic for botulinum toxin using a micro-NMR–based platform technology.

Description: Botulinum toxin is one of the most potent substances known. At present, laboratory diagnostic testing for botulism in the United States is based on the mouse bioassay that can detect as little as 0.03 ng of botulinum toxin, but requires between 1 to 2 days for results. In this project, a portable micro-nuclear magnetic resonance (?-NMR) spectrometer platform utilizing nanoparticle amplification will be developed for the rapid diagnosis of botulinum toxicity. The sensitivity of this device relies on recent advances in micro-fabrication coupled with super-paramagnetic iron oxide nanoparticle (SPION) signal amplification for NMR detection.

There have been recent advances in micro-fabrication techniques, drastically improving the sensitivity of ?-NMR, thus reducing the detection volume of NMR to below one nanoliter. In this proposal, we will be combining ?-NMR detection coils fabricated at Sandia National Laboratories with further miniaturization of permanent magnets to produce a portable ?-NMR device that can be used in diagnostic laboratories. The increase in ?-NMR sensitivity will be coupled to the relaxation-induced amplification afforded through the use of SPIONs, thereby allowing bioagent detection. The biospecificity of the SPION amplification is obtained through antibody conjugation. By utilizing different specific antibodies, it will be possible to provide rapid diagnostics of the seven distinct antigenic types of botulinum toxin. This diagnostic ?-NMR platform will be able to identify and quantify all of the Clostridium botulinum toxin types (A through G) existing in blood, serum, and tissue samples in about five minutes. This generalized ?-NMR amplification scheme can also be easily modified towards other NIAID agents, positively affecting future public health issues.