Date of Award
4-19-2010
Degree Type
Closed Dissertation
Degree Name
Doctor of Philosophy (PhD)
Department
Chemistry
First Advisor
Dr. Zhen Huang - Committee Chair
Second Advisor
Dr. Al Baumstark - Committee Member
Third Advisor
Dr. Kathy Grant - Committee Member
Fourth Advisor
Dr. Lucjan Strekowski - Committee Member
Abstract
Detection of cellular messenger RNA is a useful diagnostic strategy for the detection of patho-gens. A rapid and sensitive method for on-site detection of specific pathogens would be of great use in a number of fields. For example, a simple and inexpensive method for the detection of harmful biological agents in train stations and airports is useful for national security. Rapid detection of pathogenic E. coli strains in food production would also be of great benefit in ensuring the safety and quality of our food supply. Here we present a method for the rapid de-tection of cellular mRNA. This system is based on the 3’-labeling approach in which targeted RNA is simultaneously extended and labeled with the use of biotin labeled-dNTPs and DNA po-lymerase on an immobilized nucleic acid probe. The biotin is subsequently converted to an enzymatic label, which produces a detectable chemiluminescent reaction in the presence of substrate. Detection time of this system is short (approximately 20 minutes) because there is no need for amplification by PCR, transcription, or fluorophore labeling. This novel methodology has been successfully demonstrated by selective detection of lac Z mRNA in a total RNA sample from E. coli.
DOI
https://doi.org/10.57709/1332037
Recommended Citation
Spencer, Sarah M., "Development of RNA Microchip for the Detection of Pathogens." Dissertation, Georgia State University, 2010.
doi: https://doi.org/10.57709/1332037