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Journal of Clinical Microbiology, December 2004, p. 5528-5536, Vol. 42, No. 12
0095-1137/04/$08.00+0     DOI: 10.1128/JCM.42.12.5528-5536.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Genotypic and Phenotypic Heterogeneity among Mycobacterium tuberculosis Isolates from Pulmonary Tuberculosis Patients

Isdore Chola Shamputa,1* Leen Rigouts,1 Lovet Achale Eyongeta,1 Nabil Abdullah El Aila,1 Armand van Deun,1 Abdul Hamid Salim,2 Eve Willery,3 Camille Locht,3 Philip Supply,3 and Françoise Portaels1

Mycobacteriology Unit, Prince Leopold Institute of Tropical Medicine, Antwerp, Belgium,1 Damien Foundation Bangladesh, Banani, Dhaka, Bangladesh,2 Laboratoire des Mécanismes Moléculaires de la Pathogenèse Bactérienne, INSERM U629, Institut Pasteur de Lille, Lille, France3

Received 6 May 2004/ Returned for modification 16 June 2004/ Accepted 20 August 2004

Although the heterogeneity of Mycobacterium tuberculosis populations and the existence of mixed infections are now generally accepted, systematic studies on their relative importance are rare. In the present study, 10 individual colonies of each M. tuberculosis isolate (primary isolate) from 97 tuberculosis patients in a primarily human immunodeficiency virus-negative population were screened for heterogeneity and detectable mixed infections by spoligotyping, IS6110-based restriction fragment length polymorphism analysis, and mycobacterial interspersed repetitive unit-variable number of tandem repeat typing. The MICs of antituberculosis drugs for colonies with divergent fingerprints were determined. Infections with different bacterial subpopulations were detected in the samples from eight patients (8.2%), and the frequency of detectable mixed infections in the study population was estimated to be 2.1%. Genotypic variations were found to be independent of the drug susceptibilities, and the various molecular markers evolved independently in most cases. The predominant strains and the primary isolates always had concordant drug susceptibility and MIC testing results. These findings have implications on the interpretation of molecular epidemiology results for patient follow-up and in transmission studies.

* Corresponding author. Mailing address: Mycobacteriology Unit, Department of Microbiology, Prince Leopold Institute of Tropical Medicine, Nationalestraat 155, B-2000 Antwerp, Belgium. Phone: 32 3 247 6336. Fax: 32 3 247 6333. E-mail: icshamputa{at}itg.be.

Journal of Clinical Microbiology, December 2004, p. 5528-5536, Vol. 42, No. 12
0095-1137/04/$08.00+0     DOI: 10.1128/JCM.42.12.5528-5536.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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