Abstract by Janice Darko
Chemistry and Biochemistry
Rapid Labeling of Model Antibiotic Resistant Bacteria DNA using Fluorimetry
Global threats to treatment of bacterial infections due to antibiotic resistance (AR) have been on the rise in recent years. Current diagnostic tests identify bacteria by using blood culture which takes more than 24 hours. This study focuses on the rapid fluorescent labeling of DNA derived from bacterial AR genes (KPC & VIM) and other model DNAs using oligreen dye (OG) and molecular beacons (MB). A NanoDrop 3300 fluorospectrometer is used to take fluorescence measurements. Linear dynamic range and labeling efficiency were dependent on the following optimized conditions: dilution factor of OG (200 fold), buffer (20 mM Tris HCl, pH 8), and heat treatment of 95 °C for 15 min. Fluorescence analysis of a target DNA with a designed MB showed signal-to-background of 10 using our buffer only and 20 using our buffer with 25% ethanol. Our results provide conditions to be used in the next phase of experiments in which amplicons of AR genes will be captured on beads in flow microchips.
Keywords: antibiotic resistance. fluorescence labeling. oligreen dye. molecular beacon