In-vitro antimycobacterial drug susceptibility testing of non-tubercular mycobacteria by tetrazolium microplate assay.
BACKGROUND
Non-tubercular mycobacteria (NTM) has not been given due attention till the recent epidemic of HIV. This has led to increasing interest of health care workers in their biology, epidemiology and drug resistance. However, timely detection and drug susceptibility profiling of NTM isolates are always difficult in resource poor settings like India. Furthermore, no standardized methodology or guidelines are available to reproduce the results with clinical concordance.


OBJECTIVE
To find an alternative and rapid method for performing the drug susceptibility assay in a resource limited settings like India, we intended to evaluate the utility of Tetrazolium microplate assay (TEMA) in comparison with proportion method for reporting the drug resistance in clinical isolates of NTM.


METHODS
A total of fifty-five NTM isolates were tested for susceptibility against Streptomycin, Rifampicin, Ethambutol, Ciprofloxacin, Ofloxacin, Azithromycin, and Clarithromycin by TEMA and the results were compared with agar proportion method (APM).


RESULTS
Of the 55 isolates, 23 (41.8%) were sensitive to all the drugs and the remaining 32 (58.2%) were resistant to at least one drug. TEMA had very good concordance with APM except with minor discrepancies. Susceptibility results were obtained in the median of 5 to 9 days by TEMA. The NTM isolates were highly sensitive against Ofloxacin (98.18% sensitive) and Ciprofloxacin (90.09% sensitive). M. mucogenicum was sensitive only to Clarithromycin and resistant to all the other drugs tested. The concordance between TEMA and APM ranged between 96.4 - 100%.


CONCLUSIONS
Tetrazolium Microplate Assay is a rapid and highly reproducible method. However, it must be performed only in tertiary level Mycobacteriology laboratories with proper bio-safety conditions.
Sankar
Manimuthu Mani
MM
Clinical Microbiology Division, Department of Laboratory Medicine, All India Institute of Medical Sciences, New Delhi, India. sankar.mm@gmail.com
Gopinath
Krishnamoorthy
K
Singla
Roopak
R
Singh
Sarman
S
D000362:Agar; D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D002939:Ciprofloxacin / Q000494:pharmacology; D017291:Clarithromycin / Q000494:pharmacology; D003470:Culture Media; D024881:Drug Resistance, Bacterial; D006801:Humans; D008826:Microbial Sensitivity Tests / Q000379:methods*; D009161:Mycobacterium / Q000145:classification* / Q000187:drug effects*; D015242:Ofloxacin / Q000494:pharmacology; D015203:Reproducibility of Results; D013045:Species Specificity; D013778:Tetrazolium Salts; D013997:Time Factors
D000900:Anti-Bacterial Agents; D003470:Culture Media; D013778:Tetrazolium Salts; D002939:Ciprofloxacin; D000362:Agar; D015242:Ofloxacin; D017291:Clarithromycin