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. 1999 Aug;37(8):2602-6.
doi: 10.1128/JCM.37.8.2602-2606.1999.

Usefulness of spoligotyping To discriminate IS6110 low-copy-number Mycobacterium tuberculosis complex strains cultured in Denmark

J Bauer  1 , A B Andersen, K Kremer, H Miörner
Affiliations

Usefulness of spoligotyping To discriminate IS6110 low-copy-number Mycobacterium tuberculosis complex strains cultured in Denmark

J Bauer et al. J Clin Microbiol. 1999 Aug.

Abstract

Mycobacterium tuberculosis complex strains cultured in Denmark have been analyzed by IS6110 restriction fragment length polymorphism (RFLP) on a routine basis from 1992 and onwards. Due to the influx of immigrants with tuberculosis, the number of strains harboring only one to five copies of IS6110 has increased steadily. Since the discriminatory power of IS6110 fingerprinting for such strains is poor, we have performed additional genotyping of all low-copy-number strains by the recently described PCR-based method known as spoligotyping. A total of 311 clinical strains were typed: 14 Mycobacterium bovis BCG, 48 M. bovis, and 249 M. tuberculosis strains. Spoligotyping correctly differentiated M. bovis and M. bovis BCG from M. tuberculosis strains, but it did not differentiate M. bovis from M. bovis BCG. All M. bovis BCG strains exhibited identical spoligotype patterns. The discriminatory power of spoligotyping of low-copy-number M. tuberculosis strains was higher than that of IS6110 fingerprinting. Based on RFLP typing solely, 83% of the low-copy-number M. tuberculosis strains were found to form part of a cluster, and 75% were found to form a cluster on the basis of spoligotyping. When the two techniques were combined, the amount of clustering decreased to 55%. The combination of these two techniques might be valuable in studying the epidemiology of M. tuberculosis strains harboring few copies of the IS6110 element.

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Figures

FIG. 1
FIG. 1
RFLP patterns and spoligotyping patterns. (A) Computer-generated, normalized image showing representative examples of the RFLP patterns of 19 IS6110 low-copy-number clusters. LC-10, LC-11, and LC-12 comprise M. bovis and M. bovis BCG only. The last lane contains reference strain MT14323. (B) Computer-generated, normalized image showing representative examples of 26 spoligotyping clusters. S-15 and S-16 comprise M. bovis and M. bovis BCG only.
FIG. 2
FIG. 2
(A) Numbers of patients in 19 low-copy-number RFLP clusters. (B) Numbers of patients in 26 spoligotyping clusters. (C) Numbers of patients in 26 combined RFLP and spoligotyping clusters.
See this image and copyright information in PMC

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