In-vitro antimycobacterial drug susceptibility testing of non-tubercular mycobacteria by tetrazolium microplate assay

被引：17
作者：
Sankar M.M. [1 ]
Gopinath K. [1 ]
Singla R. [2 ]
Singh S. [1 ]
机构：
[1] Clinical Microbiology Division, Department of Laboratory Medicine, All India Institute of Medical Sciences, New Delhi
[2] Division of Respiratory Medicine, LRS Institute of Tuberculosis and Lung Diseases, New Delhi
来源：
Annals of Clinical Microbiology and Antimicrobials | / 7卷 / 1期
关键词：
Clarithromycin; Ofloxacin; Area Under Curve; Drug Susceptibility Testing; Lowenstein Jensen;
D O I：
10.1186/1476-0711-7-15
中图分类号：
学科分类号：
摘要：
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%. Conclusion: 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. © 2008 Sankar et al; licensee BioMed Central Ltd.
引用
收藏
相关论文
共 22 条
  • [1] Horsburgh Jr. C.R., Selik R., The epidemiology of disseminated non-tubercular mycobacterial infection in the acquired immunodeficiency syndrome, Am Rev Respir Dis, 139, pp. 4-7, (1989)
  • [2] Nightingale S.D., Byrd L.T., Southern P.M., Jockusch J.D., Cal S.X., Wynne B.A., Incidence of Mycobacterium avium-intracellulare complex bacteremia in human immunodeficiency virus-positive patients, J Infect Dis, 165, 6, pp. 1082-1085, (1992)
  • [3] Contreras M.A., Cheung O.T., Sanders D.E., Goldstein R.S., Pulmonary infections with non-tubercular mycobacteria, Am Rev Respir Dis, 137, pp. 149-152, (1988)
  • [4] Collins F.M., Mycobacterial disease, immuno suppression and acquired immunodeficiency syndrome, Clin Microbiol Rev, pp. 360-377, (1989)
  • [5] Philalay J.S., Palermo C.O., Hauge K.A., Rustad T.R., Cangelosi G.A., Genes required for intrinsic multidrug resistance in Mycobacterium avium, Antimicrob Agents Chemother, 48, 9, pp. 3412-3418, (2004)
  • [6] Nasha K.A., Andini N., Zhang Y., Brown-Elliott B.A., Wallace Jr. R.J., Intrinsic macrolide resistance in rapidly growing mycobacteria, Antimicrob Agents Chemother, 50, 10, pp. 3476-3478, (2006)
  • [7] Diagnosis and treatment of disease caused by Non-tubercular mycobacteria, Am J Respir Crit Care Med, 156, pp. 1-25, (1997)
  • [8] Woodley C.L., Kilburn J.O., In vitro susceptibility of Mycobacterium avium and Mycobacterium tuberculosis strains to a sipro-piperidyl rifampicin, Am Rev Respir Dis, 126, pp. 586-587, (1982)
  • [9] Heifets L., MIC as a quantitative measurement of the susceptibility of Mycobacterium avium strains to seven antituberculosis drugs, Antimicrob Agents Chemother, 32, pp. 1131-1136, (1988)
  • [10] Steadham J.E., Stall S.K., Simmank J.L., Use of the BACTEC system for drug susceptibility testing of Mycobacterium tuberculosis, M. kansasii, and M. avium complex, Diagn Microbiol Infect Dis, 3, pp. 33-40, (1985)
123