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Journal of Clinical Microbiology, December 2000, p. 4478-4484, Vol. 38, No. 12
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Genetic Heterogeneity in Mycobacterium tuberculosis Isolates Reflected in IS6110 Restriction Fragment Length Polymorphism Patterns as Low-Intensity Bands

Annette S. de Boer,1,* Kristin Kremer,2 Martien W. Borgdorff,3 Petra E. W. de Haas,2 Herre F. Heersma,4 and Dick van Soolingen2

Department of Infectious Disease Epidemiology,1 Laboratory for Infectious Diseases and Perinatal Screening, Department of Mycobacteriology,2 and Staff Bureau for Informatics and Methodological Advice,4 National Institute of Public Health and the Environment (RIVM), 3720 BA Bilthoven, and Royal Netherlands Tuberculosis Association (KNCV), 2501 CC The Hague,3 The Netherlands

Received 10 April 2000/Returned for modification 28 July 2000/Accepted 24 September 2000

Mycobacterium tuberculosis isolates with identical IS6110 restriction fragment length polymorphism (RFLP) patterns are considered to originate from the same ancestral strain and thus to reflect ongoing transmission. In this study, we investigated 1,277 IS6110 RFLP patterns for the presence of multiple low-intensity bands (LIBs), which may indicate infections with multiple M. tuberculosis strains. We did not find any multiple LIBs, suggesting that multiple infections are rare in the Netherlands. However, we did observe a few LIBs in 94 patterns (7.4%) and examined the nature of this phenomenon. With single-colony cultures it was found that LIBs mostly represent mixed bacterial populations with slightly different RFLP patterns. Mixtures were expressed in RFLP patterns as LIBs when 10 to 30% of the DNA analyzed originated from a bacterial population with another RFLP pattern. Presumably, a part of the LIBs did not represent mixed bacterial populations, as in some clusters all strains exhibited LIBs in their RFLP patterns. The occurrence of LIBs was associated with increased age in patients. This may reflect either a gradual change of the bacterial population in the human body over time or IS6110-mediated genetic adaptation of M. tuberculosis to changes in the environmental conditions during the dormant state or reactivation thereafter.

* Corresponding author. Mailing address: Department of Infectious Disease Epidemiology, National Institute of Public Health and the Environment (RIVM), P.O. Box 1, 3720 BA Bilthoven, The Netherlands. Phone: 31 30 274 3691. Fax: 31 30 274 4409. E-mail: Annette.de.Boer{at}rivm.nl.

Journal of Clinical Microbiology, December 2000, p. 4478-4484, Vol. 38, No. 12
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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