Revisiting the susceptibility testing of Mycobacterium tuberculosis to ethionamide in solid culture medium.

BACKGROUND & OBJECTIVES: Increase in the isolation of drug resistant phenotypes of Mycobacterium tuberculosis necessitates accuracy in the testing methodology. Critical concentration defining resistance for ethionamide (ETO), needs re-evaluation in accordance with the current scenario. Thus, re-evaluation of conventional minimum inhibitory concentration (MIC) and proportion sensitivity testing (PST) methods for ETO was done to identify the ideal breakpoint concentration defining resistance. METHODS: Isolates of M. tuberculosis (n=235) from new and treated patients were subjected to conventional MIC and PST methods for ETO following standard operating procedures. RESULTS: With breakpoint concentration set at 114 and 156 µg/ml, an increase in specificity was observed whereas sensitivity was high with 80 µg/ml as breakpoint concentration. Errors due to false resistant and susceptible isolates were least at 80 µg/ml concentration. INTERPRETATION & CONCLUSIONS: Performance parameters at 80 µg/ml breakpoint concentration indicated significant association between PST and MIC methods.

Stratifying low level Isoniazid resistance using additional intermediate drug concentration.

Isoniazid (INH) susceptibility testing for 100 Mycobacterium tuberculosis performed by conventional minimum inhibitory concentration (MIC) method was stratified using additional drug concentrations. Introduction of additional drug concentrations did not greatly improve the discriminatory capacity, but can be used in specialized studies pertaining to cross resistance between structural analogues of INH.

Modification of proportion sensitivity testing method for ethionamide.

Standardized methodology for drug susceptibility testing of second line drugs is vital for treatment of multi/extensively drug resistant tuberculosis. Discrepancy between laboratory methods and clinical interpretation is well established for bacteriostatic drugs such as ethionamide. Optimization of the standard proportion sensitivity testing (PST) method for ethionamide was under taken in 235 Mycobacterium tuberculosis isolates from new and previously treated pulmonary tuberculosis patients. An additional higher concentration of 57 µg/ml was evaluated against at the standard 40 µg/ml concentration in PST method. Performance parameters and agreement between the two drug concentrations was higher indicating the efficiency of PST method at its present format at 40 µg/ml and additional higher concentration of 57 µg/ml as an alternative when required.

Optimization of the conventional minimum inhibitory concentration method for drug susceptibility testing of ethionamide.

Evaluation of newer methods and optimization of existing methods for the susceptibility testing of second-line drugs, especially ethionamide, are essential when treatment of multidrug-resistant tuberculosis (MDR-TB) is warranted. The ideal method must clearly demarcate sensitive from resistant strains. Hence, optimization of the conventional minimum inhibitory concentration (MIC) method was attempted using diluted inoculum. The optimized MIC method was evaluated using 206 Mycobacterium tuberculosis strains isolated from new and previously treated tuberculosis patients and were compared with the conventional MIC method and proportion sensitivity (PST) method. The sensitivity and specificity of the optimized MIC method in comparison with the PST method was 74% and 90%. Assessment of the optimized MIC method with the conventional MIC method gave a sensitivity of and specificity of 73% and 98%. Overall agreement between the methods was found to be ⩾80%. Endowed with the ability to identify the resistant strains precisely, the optimized MIC method can be used for screening resistance to ethionamide.