|Year : 2010 | Volume
| Issue : 1 | Page : 15-19
Mycobacterium tuberculosis drug-resistance in previously treated patients in Ouagadougou, Burkina Faso
L Sangare1, S Diande2, S Kouanda3, BI Dingtoumda4, A Mourfou4, F Ouedraogo4, I Sawadogo4, B Nebie4, A Gueye4, LT Sawadogo5, AS Traore4
1 University Hospital Centre Yalgado Ouedraogo, 03 BP 7022 Ouagadougou 03; and Faculty of Health Sciences, University of Ouagadougou, 03 BP 7021 Ouagadougou 03, Burkina Faso
2 Faculty of Earth and Life Sciences, University of Ouagadougou, 03 BP 7021 Ouagadougou 03; and National Centre against Tuberculosis, Ouagadougou, Burkina Faso
3 Faculty of Health Sciences, University of Ouagadougou, 03 BP 7021 Ouagadougou 03, Burkina Faso
4 Faculty of Earth and Life Sciences, University of Ouagadougou, 03 BP 7021 Ouagadougou 03, Burkina Faso
5 National Centre against Tuberculosis, Ouagadougou, Burkina Faso
|Date of Web Publication||20-Apr-2010|
Department of Bacteriology and Virology, University Hospital Centre Yalgado Ouedraogo, 03 BP 7022 Ouagadougou 03,
Source of Support: “Secure The Future” from BMS Foundation,, Conflict of Interest: None
| Abstract|| |
Background: Tuberculosis drug-resistance becomes common in sub-Saharan Africa; however, very few data are available in Burkina Faso. The aim of this study is to assess the acquired resistance of Mycobacterium tuberculosis complex strains identified in TB patients to four first-line drugs in Ouagadougou.
Methods: One hundred and ten (110) pulmonary tuberculosis patients with acid-fast bacilli-positive sputum and in situation of failure, relapse, or treatment abandonment were included in the study. Ninety six strains, including 92 (95.8%) M. tuberculosis and 4 (4.2%) M. africanum, were isolated from the sputum samples of these patients. Their drug susceptibility testing was performed using the proportion method. The first-line drugs tested were isoniazid (INH), streptomycin (STR), ethambutol (EMB), and rifampicin (RIF).
Results: The overall drug-resistance rate of M. tuberculosis was 67.4% (n=60), including 3.4% to one drug, 18% to two, 10.1% to three, and 35.9% to four drugs. The resistance to INH, RIF, EMB, and STR were 67.4%, 51.7%, 50.6%, and 44.9%, respectively. Two strains of M. africanum were resistant to all drugs. Forty-six (51.7%) strains were multidrug-resistant (resistant to at least INH and RIF).
Conclusions: In previously treated patients, the level of resistance of M. tuberculosis complex to commonly used anti-tuberculosis drugs is very high in Ouagadougou. Our results showed that multidrug-resistant tuberculosis could be a public health problem in Burkina Faso.
| Abstract in French|| |
Arriθre-plan: Tuberculose pharmacorιsistance devient commun en Afrique; Toutefois, trθs peu de donnιes est disponibles au Burkina Faso. L'objectif de cette ιtude est pour ιvaluer la rιsistance acquise de Mycobacterium tuberculosis complexes souches identifiιes dans les tuberculeux aux quatre premiers mιdicaments de ligne ΰ Ouagadougou.
Mιthodes: Une centaine et dix patients de tuberculose pulmonaire avec acid-fast bacilles-positif expectorations et en situation de dιfaillance, abandon de rechute ou de traitement ont ιtι inclus dans l'ιtude. Quatre-vingt seize souches, y compris 92 (95.8%) M. tuberculose et 4 (4.2%) M. africanum, ont ιtι isolιes ΰ partir des ιchantillons de salive de ces patients. Leurs mιdicaments. test de sensibilitι a ιtι effectuιe ΰ l'aide de la mιthode de proportion. Le premier ligne de mιdicaments testιs ont ιtι isoniazide (INH), streptomycine (STR), ethambutol (EMB) et rifampicine (RIF).
Rιsultats: Le le taux global de la rιsistance aux mιdicaments de la tuberculose M. ιtait 67.4% (n=60), y compris de 3.4% ΰ un mιdicament, 18% ΰ deux, 10.1% ΰ trois et 35.9% aux quatre mιdicaments. La rιsistance ΰ l'INH, RIF, ECE et STR ιtait 67.4%, 51.7%, 50.6% et 44.9%, respectivement. Deux souches de M. africanum ont ιtι rιsistant ΰ toutes les drogues. Quarante-six (51.7%) souches ont ιtι tuberculoses rιsistants (rιsistant au moins INH et RIF).
Conclusions: Chez les patients traitιs prιcιdemment, le niveau de rιsistance de M . la tuberculose complexe aux mιdicaments antituberculeux couramment utilisιs est trθs ιlevιe ΰ Ouagadougou. Nos rιsultats montrent que tuberculoses multirιsistantes pourrait κtre un problθme de santι publique au Burkina Faso.
Mots clιs: La tuberculose, la rιsistance aux mιdicaments, Ouagadougou, Burkina Faso
Keywords: Burkina faso, drug resistance, Ouagadougou, tuberculosis
|How to cite this article:|
Sangare L, Diande S, Kouanda S, Dingtoumda B I, Mourfou A, Ouedraogo F, Sawadogo I, Nebie B, Gueye A, Sawadogo L T, Traore A S. Mycobacterium tuberculosis drug-resistance in previously treated patients in Ouagadougou, Burkina Faso. Ann Afr Med 2010;9:15-9
|How to cite this URL:|
Sangare L, Diande S, Kouanda S, Dingtoumda B I, Mourfou A, Ouedraogo F, Sawadogo I, Nebie B, Gueye A, Sawadogo L T, Traore A S. Mycobacterium tuberculosis drug-resistance in previously treated patients in Ouagadougou, Burkina Faso. Ann Afr Med [serial online] 2010 [cited 2019 Aug 17];9:15-9. Available from: http://www.annalsafrmed.org/text.asp?2010/9/1/15/62619
| Introduction|| |
The resurgence of tuberculosis (TB) is a major concern at the global level. The World Health Organization (WHO) estimates that 1.7 to 2 billion people are infected with Mycobacterium tuberculosis and that each year, 7-9 million develop TB of which 3 million die.  In Africa, TB prevalence remains high due mainly to the epidemic of HIV infection and the spread of multidrug-resistant (MDR) tuberculosis. ,,,,
The drug-susceptibility testing of M. tuberculosis remains a significant part of the monitoring process of the TB resistance to drugs.  Surveillance of drug-resistance, notably by periodic assessments, can guide the decision-maker in defining the standardized regimens and for assessing its quality on the field. Thus, the level and trend of resistance enables appropriate adjustments to be made either at the organizational level or changing the treatment regimens at patients' level.
In Africa, drug-susceptibility testing for M. tuberculosis is rarely done. Nevertheless, some useful information has been obtained in some countries in the course of investigations conducted by WHO and the International Union against Tuberculosis and Lung Disease (IUATLD),  or as part of studies done locally. ,,,, For the specific case of Burkina Faso, one study conducted at the Centre Muraz in Bobo-Dioulasso in 1996, a year after the adoption of directly observed treatment (DOT) in the country, showed an 8.6% overall prevalence of resistance to TB drugs in new cases.  Since then, no other study or evaluation has been conducted despite the fact that clinical treatment failures which suggest the existence of drug-resistant strains are recorded increasingly. Such a gap in information could complicate the implementation of any successful drug control program.
The aim of this study was to assess the prevalence of M. tuberculosis resistance to antibiotics in patients experiencing failure, relapse, and treatment abandonment in order to assess the quality of treatment provided in Burkina Faso.
| Materials and Methods|| |
The study was carried out in the laboratory of the National Tuberculosis Centre (NTC) in Ouagadougou, Burkina Faso. All bacteriological examinations were run in this laboratory where new TB cases are detected and where the bacteriological response to therapy of TB patients is monitored. It strongly supports the activities of the National Tuberculosis Programme (NTP).
Patients and ethical consideration
From April 2005 to February 2006, 110 patients experiencing failures, relapses, or treatment abandonment for at least 1 month and with acid fast bacilli (AFB) positive sputum smears were consecutively enrolled in a cross-sectional study. The aim of the study was explained to each participant in his language of communication. After obtaining informed consent, a standard questionnaire was completed for each patient to collect demographic data and the history of the disease such as: previous anti-TB treatment, treatment duration with or without injection (streptomycin), recognition of the anti-TB drugs on behalf of the patient, previous pulmonary radiographs and examinations of sputa.
Microscopic examination and strains isolation.
Sputum smears were prepared and stained by the Ziehl-Neelsen hot method, as recommended by IUATLD.  One AFB-positive sample from each patient was used for the culture.
The sputum samples were decontaminated with 4% NaOH, according to the Petroff method, , centrifuged, and the sediments transferred onto Loewenstein-Jensen (LJ) media, LJ supplemented with pyruvate (LJ+pyruvate), and LJ containing 5 mg/L hydrazide of the thiophene-2-carboxylic acid (TCH) (LJ+TCH). These culture media were incubated at 37°C and observed on days 3 and 7 to detect contaminations and/or fast growth of atypical mycobacteria and subsequently every week to note the growth rate and the morphology of the colonies.
Identification and drug-susceptibility test
Isolates were identified by acid fast staining, colony growth time, resistance to TCH, culture abundance in LJ+pyruvate media, and activity to usual biochemical assays (niacin, nitrate reductase, catalase activity at 22°C and 70°C). 
The antibiotics were tested according to the method of Canetti, Rist, and Grosset.  The concentrations used were 0.2 μg/mL for isoniazid (INH), 40 μg/mL for rifampicin (RIF), 4 μg/mL for streptomycin (STR), and 2 μg/mL for ethambutol (EMB). A strain was declared resistant to an antibiotic if the bacterial growth on medium with the drug was ≥1% compared to the control or sensitive when their growth was <1%. Drug resistance was acquired if the patient was previously treated with antituberculosis drugs.
A M. tuberculosis H37 ATCC 27294 strain was used for quality control in DST. Proficiency testing for culture and identification were done in collaboration with the National reference Center for Mycobacteria in Borstel (Parkallee Borstel Germany): 10 strains of M. tuberculosis Complex and 10 of atypical mycobacteria strains were used for this control.
The data were recorded and analyzed on SPSS version 12.0. The results were interpreted using standard chi-square test (χ2 ). A significant threshold was P<0.05.
| Results|| |
Demographic characteristics of patients
One hundred and ten previously treated patients were included in the study, with 73 (66.5%) cases of failure, 25 (22.7%) relapses, and 12 (10.9%) cases of treatment abandonment at least for 1 month. The failures occurred at the fifth month in 24 (21.8%) patients, at 7/8th month in 7 (7.3%) patients, and in 41 (37.3%) failures after two courses of treatment (the chronics). The age and gender distributions are presented in [Figure 1]. The sex ratio was 2.4, with 74 (67.3%) men and 36 (32.7%) women. The age range was 14-76 years old, with a mean of 38.3 11.4 years.
Mycobacterium resistance to the drugs tested
All the 110 patients were AFB-positive, while the culture was positive for 105 (95.5%), negative for 1 (0.9%), and contaminated for 4 (3.6%) patients. Among the 105 strains identified, there were 92 (87.6%) M. tuberculosis, 4 (3.8%) M. africanum, and 9 (8.6%) non-tuberculosis Mycobacterium which were excluded.
Three isolates of M. tuberculosis were excluded because of insufficient number of colonies. Drug-susceptibility testing to INH, RIF, EMB, and STR was performed with 89 M. tuberculosis and 4 M. africanum strains. The results showed that 29 (32.6%) M. tuberculosis strains were sensitive to all antibiotics tested and 60 (67.4%) were resistant. Monodrug-resistance was 67.4% to INH, 51.6% to RIF, 44.9% to STR, and 50.5% to EMB. However, 32 (35.9%) strains were MDR for INH and RIF. The other resistance details are presented in [Table 1]. Two of the four M. africanum isolates were resistant to all antibiotics tested.
Resistance of M. tuberculosis to INH, RIF, EMB, and STR separately or combined was higher in chronic cases than among cases of relapse and treatment abandonment [Table 2]. Chronic TB patients were more resistant to RIF and STR than those with failures at the 5th and 7/8th months (P=0.031 and 0.042, respectively) and also more resistant to any drug than the cases with relapse and abandonment of less than 1 month (P=0.0001).
| Discussion|| |
In this study and generally in sub-Saharan Africa, TB is more prevalent in patients aged 20 to 45 years, with a clear male prevalence.  In fact, young adults, and especially male adults, are the most economically productive and are committed in various activities from where the transmission of the tuberculosis bacillus can occur easily.  These reasons could also explain why there is more failure and relapse in this group.
The prevalence of M. tuberculosis drug-resistance in this study was 67.4%. Lower rates have been described in Nepal, Italy, Estonia,  in Mexico,  and in Prague  (40.9%, 47.2%, 58.1%, 65%, and 52%, respectively). Higher rates were found in Egypt (68.2%), Russian Federation (73.3%), Kazakhstan (82.1%),  Ivory Coast (79.0%),  and Japan (80.0%). 
The drug-resistance rate was higher in chronic cases (failure after two courses of treatment) than in other patients (failure after 5th or 7/8 th month and the relapses). It seems that the proportion of patients with resistant bacilli becomes dominant after failure of two treatment courses. It is probable that the provision of a second course of treatment as recommended by WHO will fail in the treatment of these chronic cases because when a strain is resistant to both INH and RIF, the chance of successful therapy is low. 
The chronic cases could have been detected if the culture and drug susceptibility tests were performed after the failure of the first course of treatment. Unfortunately, in vitro surveillance of M. tuberculosis resistance to drugs is not effective in the country, despite the current need. Presently, a further study is ongoing to detect XDR tuberculosis in these strains.
The MDR observed among patients in a situation of failure (particularly the chronics), relapse, and abandonment of treatment may reflect the quality of care or their poor compliance. This is an important public health problem considering the high level of resistance to drug used in the country. Monitoring of drug-resistant M. tuberculosis should be enhanced by periodic surveys to assess trends in resistance and take corrective action when necessary.
| Acknowledgments|| |
We express our gratitude to "Secure The Future" from BMS Foundation for the entire financial support. We thank Dr. Elvira Richter of National Reference Center for Mycobacteria (Parkallee Borstel Germany) for his comments, the health professionals at the CNLAT in Ouagadougou, and the laboratory of mycobacteria of the Centre Muraz in Bobo-Dioulasso for their technical assistance.
| References|| |
|1.||Dye C. Global epidemiology of tuberculosis. Lancet 2006;367:938-40. |
|2.||Center for Disease Control. Primary resistance to antituberculosis drugs, United States. MMWR Morb Mortal Wkly Rep 1983;32:521-3. |
|3.||Frieden TR, Sterling T, Pablos-Mendez A, Kilburn JO, Cauthen GM, Dooley SW. The emergence of drug-resistant tuberculosis in New York City. N Engl J Med 1993;328:521-32. |
|4.||Bloch AB, Cauthen GM, Onorato IM, Dansbury KG, Kelly GD, Driver CR, et al. Nationwide survey of drug-resistant tuberculosis in the United States. JAMA 1994;271:665-71. |
|5.||Grosset J. Frιquence et gravitι actuelles de la rιsistance de Mycobacterium tuberculosis aux antibiotiques. Ann Inst Pasteur 1993;4:196-202. |
|6.||The WHO/IUATLD Global Project on Anti-Tuberculosis Drug Resistance Surveillance 1994-1997. Anti-tuberculosis drug resistance in the world. Geneva: WHO Tuberculosis Global Programme, 1997. |
|7.||WHO/IUATLD. Anti-tuberculosis drug resistance in the world: Third global report/the WHO/IUATLD Global Project on anti-tuberculosis drug resistance surveillance 1999-2002. |
|8.||Dosso M, Bonard D, Msellati P, Bamba A, Doulhourou C, Vincent V, et al. Primary resistance to antituberculosis drugs: A national survey conducted in Cτte d'Ivoire. Int J Tuber Lung Dis 1999;3:805-9. |
|9.||Kuaban C, Bercion R, Jifon G, Cunin P, Blackett KN. Acquired anti-tuberculosis drug resistance in Yaounde, Cameroon. Int J Tuberc Lung Dis 2000;4:427-32. |
|10.||Warndorff DK, Yates M, Ngwira B, Chagaluka S, Jenkins PA, Drobniewski F, et al. Trends in antituberculosis drug resistance in Karonga, Malawi, 1986-1998. Int J Tuber Lung Dis 2000;4:752-7. |
|11.||Demissie M, Gebeyehu M, Berhane Y. Primary resistance to antituberculosis drugs in Addis Ababa, Ethiopia. Int J Tuberc Lung Dis 1997;1:64-7. |
|12.||Owusu-Dabo E, Adjei O, Meyer CG, Horstmann RD, Enimil A, Kruppa TF, et al . Mycobacterium tuberculosi s drug resistance, Ghana. Emerg Inf Dis 2006;12:1171-2. |
|13.||Ledru S, Cauchoix B, Yamιogo M, Zoubga A, Lamandι-Chiron J, Portaels F, et al. Impact of short-course therapy on tuberculosis drug resistance in South-West Burkina Faso. Tuberc Lung Dis 1996;77:429-36. |
|14.||Enarson DA, Rieder HL, Arnadottir T. Managements of tuberculosis: Guide for low income countries. 5 th ed. Paris: WHO/IUATLD; 2000. |
|15.||Petroff SA. A new and rapid method for the isolation and cultivation of tubercle bacilli directly from the sputum and feces. J Exp Med 1915;21:38-42. |
|16.||Keilty RA. A final report on the cultivation of the tubercle bacillus from the sputum by the method of Petroff. J Exp Med 1916;24:41-8. |
|17.||El Helali N, Vergez P. Identification des mycobactιries. Feuillets de Biologie 1993;34:5-19. |
|18.||Canetti G, Rist N, Grosset J. Mesure de la sensibilitι du bacille tuberculeux aux drogues antibacillaires par la mιthode des proportions. Rev Tuberc Pneumol 1963;27:217-72. |
|19.||Kouassi B, Horo K, N'douba KA, Koffi N, Ngom A, Aka-Danguy E, et al. Profil ιpidιmiologique et microbiologique des malades tuberculeux en situation d'ιchec ou de rechute ΰ Abidjan. Bull Soc Exot 2004;97:336-7. |
|20.||Ramaswamy SV, Dou SJ, Rendon A, Yang Z, Cave MD, Graviss EA.Genotypic analysis of multidrug- resistant Mycobacterium tuberculosis isolates from Monterrey, Mexico. J Med Microbial 2004;53:107-13. |
|21.||Saribas S. Resistance problems in Mycobacterium tuberculosis: Evaluation of the resistance of 166 M. tuberculosis strains against four major drugs. Euro Soc Clin Microbiol Infect Dis 2004;902:1287. |
|22.||Quy HT, Lan NT, Borgdorff MW, Grosset J, Linh PD, Tung LB, et al. Drug resistance among failure and relapse cases of tuberculosis: Is the standard re-treatment regimen adequate? In J Tuberc Lung Dis 2003;7:631-6. |
|23.||Senneville E. Traitement des patients porteurs de souches rιsistantes. Med Mal Infect 1995;25:369-76. |
[Table 1], [Table 2]
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