LDR Generated molecular beacon for rapid detection of pathogenic bacteria on microchip at single-molecule level

ANYL 167

Zhiyong Peng and Steven A. Soper. Department of Chemistry, Louisiana State Universy, Room 232 Choppin Hall, Baton Rouge, LA 70802
Detection of pathogenic bacteria and viruses are of increasing importance due to threat of bioterrorism. In this paper we describe an assay that can rapidly detect the presence of genomic DNA from certain strains of pathogens without PCR. A pair of strain specific primers (common primer and discriminating primer) were designed, which were end-labeled with Cy5 and Cy5.5, respectively. In a successful ligase detection reaction (LDR), a molecular beacon is formed by joining the two adjacent primers covalently. The hairpin structure of the molecular beacon is thermodynamically favored at the detection temperature. The fluorescence resonance energy transfer (FRET) signal from a single molecular beacon was detected in our single molecule detection experiment setup. The LDR was performed on PMMA microchip. Denaturing, ligation and detection were carried out sequentially at different temperature zones without need of thermal cycling. The pathogens present in the sample solution can be detected within 10 minutes.
 

General Papers
7:00 PM-9:00 PM, Sunday, 10 September 2006 Moscone Center -- Hall D, Poster

Sci-Mix
8:00 PM-10:00 PM, Monday, 11 September 2006 Moscone Center -- Hall D, Sci-Mix

Division of Analytical Chemistry

The 232nd ACS National Meeting, San Francisco, CA, September 10-14, 2006