Second Line Injectable Drug Resistance and Associated Genetic Mutations in Newly Diagnosed Cases of Multidrug-Resistant Tuberculosis.

Aim: To investigate the phenotypic and genotypic profile of multidrug-resistant (MDR) Mycobacterium tuberculosis (MTB) clinical isolates with reference to second-line injectable drugs (SLIDs).

Methods

A total of 110 MTB isolates, recovered consecutively from confirmed MDR-TB patients between March and June 2016, were included in this study. Phenotypic drug susceptibility testing against SLIDs (Kanamycin, Amikacin, and Capreomycin) and Ofloxacin (OFX) was performed using the MGIT 960 system. For genotypic analysis, SLID/(s) resistant (n = 13) and susceptible isolates (n = 26) were subjected to PCR and DNA sequencing for rrs, eis (promoter region), and tlyA loci of MTB. Furthermore, the identified genetic mutations were analyzed with respect to its significance in detecting phenotypic resistance. Result: Among the 110 analyzed isolates, phenotypic resistance to OFX, SLIDs, and to both was 59.1%, 11.8%, and 10.0%, respectively. Out of a total 13 SLID/(s) resistant isolates, 10 had mutations (including two novel mutations) in one or more of the targeted genes. Only one SLID susceptible MTB isolate showed mutation in the targeted region. In SLID resistant isolates, most frequent mutation detected was C-12T under eis promoter region (46.1%).

Conclusion

Mutations in rrs, eis, and tly A loci together are important in predicting SLID resistance in MTB isolates. Future molecular epidemiology studies are needed to have more insight into frequency and clinical relevance of novel mutations identified in this study.