Rapid Detection of Isoniazid Resistance in Mycobacterium tuberculosis Isolates by Multiplex Allelic Specific PCR

Abstract

Mycobacterium tuberculosis is a type of bacteria causing infectious desease called tuberculosis. According to the Makassar City Health Office, total tuberculosis (TB) patients in 2019 were 5,412 with 83% cure rate. The number of TB cases had dropped to 3,250 in 2020 with 85% cure rate before it increased again to 3,911 in 2021. This study aimed at identifying Mycobacterium TB resistance to INH (isonicotinyl hydrazide) specifically in probable tuberculosis case in Makassar. Patients with suspected tuberculosis who have not taken medication were selected as study subject. They were examined through several stages including sample decontamination, microscopic examination (ZN Method), isolation on LJ media, culture on liquid medium MGIT (Mycobacteria Growth Indicator Tube), test on.DST (Drug Susceptible Test) MGIT, and DNA extraction using specific multiple allelic (MAS) techniques. According to the results of the culture, every sample (100%) was positive. With the use of the MGIT approach, we followed up on the encouraging culture results. In this instance, we used two MGIT tubes for each sample, one for the isoniazid antibiotic control and one for the subsequent sample. From the third day following vaccination through the twelfth, observations were made. On the third day, if the control tube is positive, then the second tube is observed until the fifth day (observation of the tube containing antibiotics is carried out two days after the control tube is positive. We also use isolates from positive culture results for Multiplex Allelic Specific PCR (MAS-PCR) ).