(240c) Rapid and Sensitive Detection of Drug Resistant Tuberculosis Mutations

Multidrug resistant tuberculosis (TB) is a major health problem in both developed and developing countries. Mutations in TB bacterial genes such as rpoB, inhA and katG lead to TB drug resistance. Identification of mutations in the rpoB, inhA and katG genes of M. tuberculosis in an accurate, rapid, and inexpensive manner are very important for improving human health. Single strand conformation polymorphism (SSCP) is a simple, rapid and inexpensive method for genetic mutation detection. In order for this technique to be clinically feasible, it must be rapid, automated, and sensitive. In this work, we focused on optimizing capillary electrophoresis (CE)-SSCP, we have investigated the effects of important variables including the polymer separation matrix and the capillary wall coating to meet the high resolution requirements for a method to be used in routine clinical practice. It was observed that 4.5% (w/v) PDMA is the optimum polymer concentration including its ability to act as a coating for the capillary walls. We demonstrated the ability of CE-SSCP to detect TB drug resistance single-base mutations in the rpoB and inhA-mabA regulatory region genes in less than one hour, even when mixed with other mycobacteria.