ULTRA-FAST PATHOGEN TOXIN DETECTION ASSAY BASED ON MICROWAVE-ACCELERATED METAL-ENHANCED FLUORESCENCE

摘要

The present invention provides for a system and method to detect low levels of the anthrax protective antigen (PA) exotoxin in biological fluids, wherein the system uses a metal-enhanced fluorescence (MEF)-PA assay in combination with microwave-accelerated PA protein surface absorption. Microwave irradiation rapidly accelerates PA deposition onto the surface adjacent to deposited metallic particles and significantly speeding up the MEF-PA assay and resulting in a total assay run time of less than 40 min with an analytical sensitivity of less than 1 pg/ml PA.

主权项

1. A system for detecting a protective antigenic protein of a pathogenic microorganism in a testing sample suspected of including same, the system comprising:
a) a substrate surface adapted for binding of the protective antigenic protein; b) metallic particles deposited on the substrate surface, wherein the metallic particles have a diameter from about 40 nm to about 120 nm and separated from each other with a distance from about 10 nm to 50 nm; c) a first antibody having binding affinity for the protective antigen protein; d) a second antibody have binding affinity for the first antibody, wherein the second antibody comprises an at least one excitable molecule, wherein the excitable molecule is selected from the group of an intrinsic fluorophore, extrinsic fluorophore, and fluorescent dye, upon binding to the first antibody positions the excitable molecule from about 5 nm to about 30 nm from at least one adjacent metallic particles; e) a source of electromagnetic energy to deliver radiation in a range of UV to IR and in an amount sufficient to excite the excitable molecule; f) a source of microwave energy in an amount sufficient to increase binding reactions in the system; and g) a detector to measure emissions from the excitable molecule and/or metallic particles.