University of South Florida

Daniel Van Lim

Dawn M Hunter

Edward Turos

Peter George Stroot

Rudiger Schlaf

Kelly M Leach

Anthony J Cascio

Edward T Carpenter

Sonja S Dickey

Timothy Long

Joyce Marie Stroot

Marianne F Brannon

The subject invention concerns materials and methods for detecting a target cell in a population. Methods of the invention comprise internally labeling cells via fluorescence in situ hybridization (FISH) using probes that target rRNA, followed by binding of capture antibodies targeted (CAT) for specific cell surface epitopes on the target cells. In one embodiment, the target cells are bacterial cells.

Methods and materials for capture antibody targeted fluorescent in-situ hybridization (CAT-FISH)

Disclosed is a method and associated device for the rapid identification of viable bacterial contaminants in food products. The method detects viable microbes by using a combined ATP-bioluminescence immunoassay.O157:H7 andwere selected as target organisms in various matrices including ground beef homogenate, apple juice, milk, and phosphate-buffered saline. Specific antibodies were immobilized on the surface of well plates in which the sample matrices were incubated. The plates were washed, and the wells were incubated with BacTiter-Glo reagent in Mueller-Hinton II broth. Bioluminescent output was measured with a luminometer and signal-to-noise ratios were calculated. The LOD was not affected by the presence of non-target cells. A strong linear correlation was observed between the number of cells and luminescent output over 4 orders of magnitude. This method provides a means of simultaneously detecting and identifying viable pathogens in complex matrices.

Detection plate for ATP-bioluminescence immunoassay and method of manufacturing

Disclosed is a method and associated device for the rapid identification of viable bacterial contaminants in food products. The method detects viable microbes by using a combined ATP-bioluminescence immunoassay.O157:H7 andserovar Typhimurium were selected as target organisms in various matrices including ground beef homogenate, apple juice, milk, and phosphate-buffered saline. Specific antibodies were immobilized on the surface of well plates in which the sample matrices were incubated. The plates were washed, and the wells were incubated with BacTiter-Glo reagent in Mueller-Hinton II broth. Bioluminescent output was measured with a luminometer and signal-to-noise ratios were calculated. The LOD was not affected by the presence of non-target cells. A strong linear correlation was observed between the number of cells and luminescent output over 4 orders of magnitude. This method provides a means of simultaneously detecting and identifying viable pathogens in complex matrices.

ATP-bioluminescence immunoassay

An electrospray (ES)-based deposition system enabling the coating an impervious substrate, such as a glass slide, with biological materials in a vacuum. Distilled water or a buffer is used as the solvent; no other solvents are used thereby eliminating hazardous waste from the process. Movement across differential pumping stages causes evaporation of the solvent occurs resulting in shrinkage of the remaining constituents with an increase of the charge density. The resulting ion beam enters a vacuum chamber and the beam impinges on the substrate, whereby a thin layer is deposited thereon. The spray can be focused to a specific area allowing patterning of the substrate if desired. The amount of coating can be controlled and a specified number of coats of the same or different molecules can be added to the surface.

Electrospray depositing system for biological materials

N-thiolated &bgr;-lactam antibiotics

Growing community of inventors

Tampa, FL, United States of America

Daniel Van Lim