DETECTION OF MULTIDRUG-RESISTANT MYCOBACTERIUM TUBERCULOSIS DIRECTLY FROM SPUTUM SAMPLES OF PATIENTS FROM JAKARTA, INDONESIA BY RADIOISOTOPE-BASED PCR-DOT BLOT HYBRIDIZATION

The problem of eradicating tuberculosis (TB) has become more com- plicated by the emergence of multidrug resistant TB (MDR-TB). Any rapid labo- ratory method that can be used to detect drug susceptibility of Mycobacterium tuberculosis (MTB) is urgently needed. In this study, we employed the radioisotope ( 32 P)-based PCR-dot blot hybridization method on sputum samples from patients in Jakarta, Indonesia. Bacterial DNA was extracted using BOOM method. KatG and rpo were amplified by PCR and katG315 or rpob531 mutations were identi- fied by dot blot hybridization. Of 100 samples, 11% and 22% showed presence of mutation at codons 315 (AGC ACC) of katG and 531 (TCG TTG) of rpo , respectively. Five percent of the samples showed both mutations. This method is rapid, sensitive, and reliable and can be used to screen large numbers of samples in epidemiological studies.