Farjana Rahman

Molecular Approaches for Detection of the Multi-Drug Resistant Tuberculosis (MDR-TB) in Bangladesh

The principal obstacles in the treatment of tuberculosis (TB) are delayed and inaccurate diagnosis which often leads to the onset of the drug resistant TB cases. To avail the appropriate treatment of the patients and to hinder the transmission of drug-resistant TB, accurate and rapid detection of resistant isolates is critical. Present study was designed to demonstrate the efficacy of molecular techniques inclusive of line probe assay (LPA) and GeneXpert MTB/RIF methods for the detection of multi-drug resistant (MDR) TB. Sputum samples from 300 different categories of treated and new TB cases were tested for the detection of possible mutation in the resistance specific genes (rpoB, inhA and katG) through Genotype MTBDRplus assay or LPA and GeneXpert MTB/RIF tests. Culture based conventional drug susceptibility test (DST) was also carried out to measure the efficacy of the molecular methods employed. Among 300 samples, 191 (63.7%) and 193 (64.3%) cases were found to be resistant against rifampicin in LPA and GeneXpert methods, respectively; while 189 (63%) cases of rifampicin resistance were detected by conventional DST methods. On the other hand, 196 (65.3%) and 191 (63.7%) isolates showed isoniazid resistance as detected by LPA and conventional drug susceptibility test (DST), respectively. Among the drug resistant isolates (collectively 198 in LPA and 193 in conventional DST), 189 (95.6%) and 187 (96.9%) were considered to be MDR as examined by LPA and conventional DST, respectively. Category-II and -IV patients encountered higher frequency of drug resistance compared to those from category-I and new cases. Considering the higher sensitivity, specificity and accuracy along with the required time to results significantly shorter, our study supports the adoption of LPA and GeneXpert assay as efficient tools in detecting drug resistant TB in Bangladesh.

Evaluation of the effectiveness of BACTEC MGIT 960 for the detection of mycobacteria in Bangladesh

OBJECTIVE Tuberculosis (TB) caused by Mycobacterium tuberculosis has been identified as a re-emerging infectious disease with public health importance globally. Exploitation of new laboratory techniques for precise identification of mycobacteria in clinical specimens is of great importance to improve the diagnosis as part of the global TB control efforts. METHODS The current study was conducted for the evaluation of BACTEC MGIT 960 method in comparison with Lowenstein-Jensen (LJ) culture and light emitting diode (LED) fluorescence microscopy for isolation of mycobacteria among TB suspects from Bangladesh. A total of 421 specimens were tested with these methods. RESULTS Among the tested samples, 3.6% (n=15) were LED fluorescence microscopy positive; while 18 (4.2%) and 45 (10.6%) were recovered from LJ and MGIT 960 culture. The relative positivity found through MGIT 960 system were 60% and 66.7% higher than that of LJ culture and LED fluorescence microscopy, respectively. Recovery rate of Mycobacterium tuberculosis complex ([MTC], 21 by MGIT and 16 by LJ culture) and non-tubercular mycobacteria ([NTM], 24 by MGIT and 2 by LJ culture) by MGIT 960 was 24% and 96% greater, respectively than LJ culture. Moreover, MGIT 960 was found to be highly sensitive (100%), specific (93.3%), accurate (93.6%) and a more rapid method in detecting mycobacteria when compared with LJ culture. CONCLUSION Extended recovery of NTM and MTC through MGIT 960 urged frequent application of this method to detect mycobacteria more effectively and rapidly.

Comparisons among the diagnostic methods used for the detection of extra-pulmonary tuberculosis in Bangladesh

The present study was an attempt to establish a suitable method for the effective diagnosis of extra-pulmonary tuberculosis in Bangladesh. In this regard, detection of Mycobacterium tuberculosis from 390 different extra-pulmonary specimens was performed by Bright-Field microscopy, light-emitting diode fluorescence microscopy and Lowenstein-Jensen culture methods, followed by an extensive comparison among these methods. M. tuberculosis was detected in 53 cases through the conventional Lowenstein-Jensen culture method; 49 cases were detected under Bright-Field microscope, whereas the light-emitting diode fluorescence microscopy detected 64 cases. Out of 53 culture-positive isolates, 12 were found to be multi-drug resistant. Light-emitting diode fluorescence microscopy was found to be more sensitive and effective than both the Bright-Field microscopy and the Lowenstein-Jensen culture methods. Incidentally, light-emitting diode fluorescence microscopy appeared imperative to detecting the multi-drug resistant tuberculosis.