Rapid identification and tracing the origin of pathogenic bacteria are imperative in response to a bioterrorist attack or an infectious disease outbreak. Pulsed-Field Gel Electrophoresis (PFGE) fingerprinting is frequently used for these tasks, particularly for tracing bacterium origins. However, the short fragment bands are lost in PFGE when extended separation times are used to resolve the long DNA molecules, thus reduce the detection specificity, because the lengths of the short fragments provide additional genotype information that could be critical to discriminate two similar genomes. Another drawback of PFGE is that the electrophoresis is too slow. In a rapid (24-hour) PFGE method the electrophoresis takes 14-18 hours. Additionally, the intra- and inter-laboratory reproducibility of PFGE also needs to be improved. Although diverse approaches such as entropic traps and DNA prisms have been explored for DNA separations, none have thus far been practically utilized. We have recently discovered a new and efficient technique to resolve broad size ranges of DNA molecules. In this application we propose to construct a novel instrument to demonstrate the proof-of-principle of this new technique for high-speed and accurate microbial identification.

Public Health Relevance

We have recently discovered a new and efficient technique to resolve broad size ranges of DNA molecules. On the basis of this discovery and as a first step toward our ultimate goal of developing a system for fast microbial identification, we plan to construct a novel instrument to demonstrate the proof-of-principle of this new technique for high-speed and accurate microbial identification.

Agency
National Institute of Health (NIH)
Type
Exploratory/Developmental Grants (R21)
Project #
5R21GM104526-03
Application #
8724529
Study Section
Special Emphasis Panel (ZRR1)
Program Officer
Friedman, Fred K
Project Start
Project End
Budget Start
Budget End
Support Year
3
Fiscal Year
2014
Total Cost
Indirect Cost
Name
University of Oklahoma Norman
Department
Chemistry
Type
Schools of Arts and Sciences
DUNS #
City
Norman
State
OK
Country
United States
Zip Code
73019