Volume 16, Number 2—February 2010
Mycobacterium tuberculosis Beijing Strain, Bamako, Mali
To the Editor: Mycobacterium tuberculosis has >36 identified genotype families (1). Four genotypes cause 35% of documented cases of active tuberculosis (TB): Beijing (10%–11%), Latin American–Mediterranean (9.3%), Haarlem (7.5%), and the X clade (7%) (1,2) The Beijing clade strains, reported in 1995 from the People’s Republic of China, are widely recognized as highly pathogenic with a possible predilection for multidrug resistance (3). Predominant in Asia, these strains have been documented in other parts of the world (1,4,5). The virulence, propensity to become resistant, and distinct geographic distribution of the Beijing clade suggest it may have some adaptive advantage in producing disease in humans. Limited data suggest that its presence in Africa is low (2,4,5).
In Bamako, Mali, 2 patients with active pulmonary TB came to the research clinic at Point G Hospital, affiliated with the University of Bamako Medical School, for recruitment under a US National Institute of Allergy and Infectious Diseases’ institutional review board–approved protocol. The first patient, a previously healthy 34-year-old man, sought treatment in March 2008. He had a 3-month history of fever, cough, shortness of breath, and left-sided chest pain; respiratory rate of 24/min; temperature of 36.8ºC; and pulse rate of 68/min. He weighed 60 kg. His leukocyte count was 8,700 cells/μL, and he was positive for HIV-1 with a CD4+ T-cell count of 468 cells/μL. He reported contact with persons from other countries in Africa, China, and other parts of Asia.
Chest radiograph showed a cavitary lesion on the left upper lobe and opacities throughout the left lung. Three sputum samples collected 3 days apart were digested and decontaminated with N-acetyl-L-cysteine, 4% NaOH; concentrated by high-speed centrifugation; stained with auramine-rhodamine; and evaluated by using fluorescent microscopy. The many acid-fast bacilli (AFB) seen were identified by using nucleic acid probes (AccuProbe, Gen-Probe, San Diego CA, USA). Antimycobacterial drug susceptibility was determined by using a manual indirect susceptibility test (mycobacterial growth indicator tube, AST SIRE System; BBL, Becton Dickinson, Franklin Lakes, NJ, USA) showed the isolate sensitive to isoniazid (0.1 μg/mL), rifampin (1.0 μg/mL), and ethambutol (3.5 μg/mL) but resistant to streptomycin (0.8 μg/mL). Spoligotyping using a commercially available kit (Spoligotyping Isogen Life Science, De Meern, the Netherlands) showed characteristics of the Beijing clade (Appendix Figure, panel A) (6).
The patient began treatment with the standard first-line regimen of isoniazid, rifampin, pyrazinamide, and ethambutol fixed-dose combination (Svizera Laboratory, Mumbai, India) according to Malian National Guidelines. Follow-up sputum samples at 13 and 18 weeks of treatment were smear- and culture-negative for AFB.
The second patient, a 28-year-old woman, sought treatment in July 2008. For 1 year, she had received first-line and retreatment regimens that failed to clear her sputum of AFB. She had begun second-line treatment for multidrug-resistant disease 2 days earlier. She had a history of fever, cough, and weight loss; temperature of 37.1ºC; heart rate of 104 beats/min; respiratory rate of 24/min; and blood pressure of 90/60 Hg mm. She weighed 49 kg. Leukocyte count was 9,400 cells/μL. Serologic results for HIV-1 and -2 were negative. Chest radiograph showed a right apical cavitary lesion and a fibrotic lesion in the right middle lung field. She did not recall any exposure to TB. She worked as an assistant at a local telephone center.
Two sputum samples, processed as described above, were positive for, and Gen-Probe testing confirmed, M. tuberculosis. According to antimycobacterial susceptibility testing, the strain was resistant to isoniazid (0.1 μg/mL), rifampin (1.0 μg/mL), ethambutol (3.5 μg/mL), and streptomycin (0.8 μg/mL). Spoligotyping confirmed the strain as Beijing clade, and restriction fragment length polymorphism (7) confirmed that it differed from that of patient 1 (Appendix Figure, panel B).
The relevance of different genotypes, such as the Beijing clade, to disease progression is being studied. Evidence indicates the genotype may factor in transmission or pathogenesis. In a study in Cape Town, South Africa, disease produced by the Beijing clade increased exponentially over time, suggesting a possible pathogenic advantage; although most cases were drug susceptible, the likelihood of unsuccessful treatment was greater than for non-Beijing variants (8). Although the Beijing clade does not appear to have greater propensity than non-Beijing genotypes for acquiring resistance, certain variants within the group that become multidrug resistant may be more likely to acquire such resistance. Beijing strains particularly may tend to acquire resistance more easily than others under conditions of suboptimal treatment (9). In Cape Town during 2000–2003, the Beijing clade as a cause of disease in children increased from 13% to 33%, suggesting a selective advantage in transmissibility and disease production (10).
These cases highlight the need to diagnose disease and resistance early and to begin appropriate treatment in TB-endemic countries. Knowledge of circulating strains and their resistance patterns is essential to developing effective programs to curtail the spread of TB within the country and the region; in this era of globalization, it is required for the successful control of TB worldwide.
We thank the Center de Recherche et Formation (SEREFO) team members for their support. We also thank the clinical staff, Souleymane Diallo, Sounkalo Dao, Samba Diop, Bindongo P.P. Dembele, Hamadoun Kassambara, Drissa Goita, and Ousmane M’baye for their efforts in patient recruitment; and the laboratory staff, Guindo Oumar, Djeneba Dabitao, Hama Diallo, Yeya D.S. Sarro, Mariam Tall, Nadie Coulibaly, and Bourahima Kone for patient screening and timely completion of all laboratory results. We also thank Mark Parta for help with the spoligotyping assay, Ousmane Koita for editorial help, and Christian Yoder for supporting the construction of the Biosaftey Level 3 laboratory.
This work was conducted at the Mali International Centers for Excellence in Research site funded by the National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland, USA.
- Filliol I, Driscoll JR, van Soolingen D, Kreiswirth BN, Kremer K, Valétudie G, Global distribution of Mycobacterium tuberculosis spoligotypes. Emerg Infect Dis. 2002;8:1347–9.
- European Concerted Action on New Generation Genetic Markers and Techniques for the Epidemiology and Control of Tuberculosis. Beijing/W genotype Mycobacterium tuberculosis and drug resistance. Emerg Infect Dis. 2006;12:736–43.
- van Soolingen D, Qian L, de Haas PE, Douglas JT, Traore H, Portaels F, Predominance of a single genotype of Mycobacterium tuberculosis in countries of east Asia. J Clin Microbiol. 1995;33:3234–8.
- Glynn JR, Whiteley J, Bifani PJ, Kremer K, van Soolingen D. Worldwide occurrence of Beijing/W strains of Mycobacterium tuberculosis: a systematic review. Emerg Infect Dis. 2002;8:843–9.
- Bifani PJ, Mathema B, Kurepina NE, Kreiswirth BN. Global dissemination of the Mycobacterium tuberculosis W-Beijing family strains [review]. Trends Microbiol. 2002;10:45–52.
- Kremer K, Glynn JR, Lillebaek T, Niemann S, Kurepina NE, Kreiswirth BN, Definition of the Beijing/W lineage of Mycobacterium tuberculosis on the basis of genetic markers. J Clin Microbiol. 2004;42:4040–9.
- Van Embden JD, Cave MD, Crawford JT, Dale JW, Eisenach KD, Gicquel B, Strain identification of Mycobacterium tuberculosis by DNA fingerprinting: recommendations for a standardized methodology. J Clin Microbiol. 1993;31:406–9.
- van der Spuy GD, Kremer K, Ndabambi SL, Beyers N, Dunbar R, Marais BJ, Changing Mycobacterium tuberculosis population highlights clade-specific pathogenic characteristics. Tuberculosis (Edinb). 2009;89:120–5. Epub 2008 Dec 2.
- Werngren J, Hoffner SE. Drug-susceptible Mycobacterium tuberculosis Beijing genotype does not develop mutation-conferred resistance to rifampin at an elevated rate. J Clin Microbiol. 2003;41:1520–4.
- Cowley D, Govender D, February B, Wolfe M, Steyn L, Evans J, Recent and rapid emergence of W-Beijing strains of Mycobacterium tuberculosis in Cape Town, South Africa. Clin Infect Dis. 2008;47:1252–9.