Mycobacterium tuberculosis Uganda II is more susceptible to rifampicin and isoniazid compared to Beijing and Delhi/CAS families.
BACKGROUND
Mycobacterium tuberculosis Uganda family is the predominant cause of tuberculosis in Uganda. Reasons for this are not clear but are likely to be due to the rampant person-to-person transmission or delayed susceptibility of the organism to drugs during treatment, which may lead to continuous shedding of infectious bacilli, among others. The objective of this study was to determine in vitro, the susceptibility patterns of M. tuberculosis Uganda family compared with Beijing and Delhi/CAS, other M. tuberculosis sub-lineages that also circulate in Uganda but are not as prevalent. The comparisons were made after 10 days of exposure of the strains to Rifampicin and Isoniazid, the most important first-line anti-tuberculosis drugs.


METHODS
Minimum inhibitory concentrations (MICs) for three Isoniazid- and Rifampicin-susceptible M. tuberculosis strains (Uganda II, Beijing and Delhi/CAS families) were determined by micro-dilution plate assay. Killing curves for each strain were deduced from colony forming units after exposure to Isoniazid and Rifampicin on days 0, 2, 4, 6, 8, and 10 under aerobic and oxygen-depleted conditions. Data were analyzed with GraphPad Prism 5 software.


RESULTS
The MIC for Isoniazid was 0.05 μg/ml for M. tuberculosis Uganda II, and 0.03 μg/ml for M. tuberculosis Beijing and Delhi/CAS. Rifampicin MIC was 1 μg/ml for M. tuberculosis Uganda II, and 0.12 μg/ml for Beijing and Delhi/CAS. At low Rifampicin (0.03-2.5 μg/ml) and Isoniazid (0.12-5 μg/ml) concentrations under aerobic conditions, there was no significant difference in susceptibility patterns between M. tuberculosis Uganda II and Beijing or Delhi/CAS. However, at high Rifampicin (5 μg/ml) and Isoniazid (1.25 μg/ml) concentrations under oxygen-depleted conditions, M. tuberculosis Uganda II was more susceptible to the drugs compared with Beijing or Delhi/CAS families.


CONCLUSIONS
The predominance of M. tuberculosis Uganda II family as the main causative agent of tuberculosis in Uganda is not attributed to its susceptibility behavior to Isoniazid and Rifampicin. Probably, its predominance is due to differences in the immune defenses in the general population against the strains, given that Beijing and Delhi/CAS families may have been introduced more recently. Further research beyond susceptibility to anti-tuberculosis drugs is required to fully explore tuberculosis strain predominance in Uganda.
National Tuberculosis and Leprosy Program, Kampala, Uganda.|National TB Reference Laboratory, Kampala, Uganda.
George W
GW
Kasule
Department of Immunology and Molecular Biology, College of Health Sciences, Makerere University, Kampala, Uganda.|Department of Medical Microbiology, College of Health Sciences, Makerere University, Kampala, Uganda.
David P
DP
Kateete
National Tuberculosis and Leprosy Program, Kampala, Uganda. moses.joloba@case.edu.|National TB Reference Laboratory, Kampala, Uganda. moses.joloba@case.edu.|Department of Immunology and Molecular Biology, College of Health Sciences, Makerere University, Kampala, Uganda. moses.joloba@case.edu.|Department of Medical Microbiology, College of Health Sciences, Makerere University, Kampala, Uganda. moses.joloba@case.edu.
Moses L
ML
Joloba
D005838:Genotype; D006801:Humans; D007538:Isoniazid / Q000494:pharmacology*; D008826:Microbial Sensitivity Tests; D009169:Mycobacterium tuberculosis / Q000145:classification / Q000187:drug effects* / Q000235:genetics; D010100:Oxygen / Q000378:metabolism; D012293:Rifampin / Q000494:pharmacology*; D014376:Tuberculosis / Q000175:diagnosis / Q000382:microbiology
D010100:Oxygen; D007538:Isoniazid; D012293:Rifampin
Aerobic; Drug susceptibility; Genotype; Isoniazid; Minimum inhibitory concentrations; Mycobacterium tuberculosis Uganda family; Oxygen depleted; Rifampicin; Strain; Time-killing-curves