| Tuberculosis (TB) is a major health problem in both developed and developing countries. There are almost 3 million TB deaths per year due to the long diagnosis time and problems with drug resistant TB. Mycobacterium (M.) tuberculosis, M. microti, M. africanum, M. bovis and M. bovis BCG are TB causing bacteria and mutations in genes such as rpoB, inhA and katG lead to TB drug resistance. Identification of TB causing bacteria from non-TB bacteria and identification of mutations in the rpoB, inhA, katG genes of M. tuberculosis in an accurate, rapid, and inexpensive manner are very important for improving human health.;In this project, we focused on amplification, purification and identification of different drug resistance mutations. We demonstrate the ability of our optimized technique (CE-SSCP) to detect TB drug resistance single-base mutations in the rpoB, inhA-mabA regulatory region, and katG genes in less than one hour.;Single-strand conformational 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 and automated. Therefore, we have implemented SSCP-capillary electrophoresis (CE). We have investigated the effects of variables such as polymer separation matrix concentration, capillary wall coating, electric field strength, and temperature on resolution to meet the high resolution requirements for a method to be used in routine clinical practice and observed that 4.5% PDMA is the optimum polymer, PDMA is the optimum capillary coating, 20°C is the optimum temperature and 278 V/cm is the optimum electric field strength. |
