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Journal of Clinical Microbiology, September 2002, p. 3398-3405, Vol. 40, No. 9
0095-1137/02/$04.00+0     DOI: 10.1128/JCM.40.9.3398-3405.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Mycobacterium africanum Subtype II Is Associated with Two Distinct Genotypes and Is a Major Cause of Human Tuberculosis in Kampala, Uganda

S. Niemann,1* S. Rüsch-Gerdes,1 M. L. Joloba,2 C. C. Whalen,3 D. Guwatudde,2,3 J. J. Ellner,4 K. Eisenach,5 N. Fumokong,5 J. L. Johnson,3 T. Aisu,2 R. D. Mugerwa,2 A. Okwera,2 and S. K. Schwander4

National Reference Center for Mycobacteria, Research Center Borstel, Borstel, Germany,1 Departments of Medicine and Medical Microbiology, Makerere University, Kampala, Uganda,2 Departments of Epidemiology and Biostatistics and of Medicine, Case Western Reserve University School of Medicine, Cleveland, Ohio 44106,3 Department of Medicine and Ruy V. Lourenco Center for the Study of Emerging and Reemerging Pathogens, University of Medicine and Dentistry-New Jersey Medical School, Newark, New Jersey 07103,4 Departments of Pathology and Immunology, University of Arkansas for Medical Sciences, Little Rock, Arkansas 722055

Received 31 January 2002/ Returned for modification 18 March 2002/ Accepted 14 June 2002

The population structure of 234 Mycobacterium tuberculosis complex strains obtained during 1995 and 1997 from tuberculosis patients living in Kampala, Uganda (East Africa), was analyzed by routine laboratory procedures, spoligotyping, and IS6110 restriction fragment length polymorphism (RFLP) typing. According to biochemical test results, 157 isolates (67%) were classified as M. africanum subtype II (resistant to thiophen-2-carboxylic acid hydrazide), 76 isolates (32%) were classified as M. tuberculosis, and 1 isolate was classified as classical M. bovis. Spoligotyping did not lead to clear differentiation of M. tuberculosis and M. africanum, but all M. africanum subtype II isolates lacked spacers 33 to 36, differentiating them from M. africanum subtype I. Moreover, spoligotyping was not sufficient for differentiation of isolates on the strain level, since 193 (82%) were grouped into clusters. In contrast, in the IS6110-based dendrogram, M. africanum strains were clustered into two closely related strain families (Uganda I and II) and clearly separated from the M. tuberculosis isolates. A further characteristic of both M. africanum subtype II families was the absence of spoligotype spacer 40. All strains of family I also lacked spacer 43. The clustering rate obtained by the combination of spoligotyping and RFLP IS6110 analysis was similar for M. africanum and M. tuberculosis, as 46% and 49% of the respective isolates were grouped into clusters. The results presented demonstrate that M. africanum subtype II isolates from Kampala, Uganda, belong to two closely related genotypes, which may represent unique phylogenetic branches within the M. tuberculosis complex. We conclude that M. africanum subtype II is the main cause of human tuberculosis in Kampala, Uganda.

* Corresponding author. Mailing address: National Reference Center for Mycobacteria, Forschungszentrum Borstel, Parkallee 18, D-23845 Borstel, Germany. Phone: (49) 4537 188658. Fax: (49) 4537 188311. E-mail: sniemann{at}fz-borstel.de.

Journal of Clinical Microbiology, September 2002, p. 3398-3405, Vol. 40, No. 9
0095-1137/02/$04.00+0     DOI: 10.1128/JCM.40.9.3398-3405.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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