Enhanced Optical Methods to Break Spore Coating
There is a need for rapid and accurate detection of bacterial spores such as Bacillus anthracis. Currently, two detection techniques are being used to rapidly identify bacterial spores. These techniques are MALDI_MS and PCR-amplification. These techniques identify the spores by evaluating biomarkers inside the cell. Both of these methods require spore disruption to release biological biomarkers before the identification can take place.
This invention is a novel method to open spore coatings and cell walls using light absorbing dyes and an optical light source (that corresponds to the absorption maximum of the absorbing dye) to facilitate the release of internal cell bio-molecules such as proteins, lipids, nucleic acids, etc. Spores coatings are difficult to damage because spores contain a dense protein layer comprising about 20 proteins that shields the interior of the spore from external environmental attack. Bacterial spores can survive indefinite periods of starvation, desiccation and exposure to toxic chemicals because of theis dense coating of proteins and cell wall structure. The attachment of organic dyes to the surface of the spore coating can provide an enhanced method of damaging spore coatings because organic dyes are highly absorbing at specific wavelengths of light. By matching the organic dye to the laser source, it is possible to deliver energy on the coating of the spore using a greater range of light source. Organic dyes are chosen because of their specific properties to that 1) have absorption spectra that match desired laser wavelengths, 2) that have demonstrated ability to chemically bind to cell surfaces, 3) that can be coated in water or other simple solvent system, and 4) can be irradiated to solid –phase form, i.e. no solvent required.
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