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. 1996 Oct;34(10):2469–2474. doi: 10.1128/jcm.34.10.2469-2474.1996

Differentiation of Mycobacterium bovis isolates from animals by DNA typing.

R A Skuce 1, D Brittain 1, M S Hughes 1, S D Neill 1
PMCID: PMC229296  PMID: 8880502

Abstract

The insertion sequence IS6110 and the direct repeat (DR) specific to tuberculosis complex mycobacteria and the highly repeated DNA sequence, the polymorphic GC-rich repeat sequence (PGRS), were systematically used to identify restriction fragment length polymorphisms (RFLPs) within 210 isolates of Mycobacterium bovis. The isolates were primarily of bovine origin, but isolates from badgers, feral deer, sheep, humans, and a pig were included. The RFLP probes IS6110, DR, and PGRS individually identified 17, 18, and 18 different RFLP types, respectively, but in combination these probes identified a total of 39 different M. bovis RFLP types. The recommendations (J. D. A. van Embden, M. D. Cave, J. T. Crawford, J. W. Dale, K. D. Eisenach, B. Gicquel, P. W. M. Hermans, C. Martin, R. McAdam, T. M. Shinnick, and P. M. Small, J. Clin. Microbiol. 31:406-409, 1993) for a standardized RFLP analysis for M. tuberculosis were adapted to facilitate gel documentation, image analysis, and construction of a database of RFLP types. In the present study the same M. bovis RFLP types were evident in the various animal species included, indicating that the strains were not host restricted. Application of these techniques to defined field studies should help elucidate more accurately aspects of the epidemiology of bovine tuberculosis in different countries.

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Selected References

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  1. Cheeseman C. L., Wilesmith J. W., Stuart F. A. Tuberculosis: the disease and its epidemiology in the badger, a review. Epidemiol Infect. 1989 Aug;103(1):113–125. doi: 10.1017/s0950268800030417. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Collins D. M., Erasmuson S. K., Stephens D. M., Yates G. F., De Lisle G. W. DNA fingerprinting of Mycobacterium bovis strains by restriction fragment analysis and hybridization with insertion elements IS1081 and IS6110. J Clin Microbiol. 1993 May;31(5):1143–1147. doi: 10.1128/jcm.31.5.1143-1147.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Collins D. M., Gabric D. M., de Lisle G. W. Typing of Mycobacterium bovis isolates from cattle and other animals in the same locality. N Z Vet J. 1988 Mar;36(1):45–46. doi: 10.1080/00480169.1988.35476. [DOI] [PubMed] [Google Scholar]
  4. Collins D. M., Stephens D. M. Identification of an insertion sequence, IS1081, in Mycobacterium bovis. FEMS Microbiol Lett. 1991 Sep 15;67(1):11–15. doi: 10.1016/0378-1097(91)90435-d. [DOI] [PubMed] [Google Scholar]
  5. Collins D. M., de Lisle G. W., Collins J. D., Costello E. DNA restriction fragment typing of Mycobacterium bovis isolates from cattle and badgers in Ireland. Vet Rec. 1994 Jun 25;134(26):681–682. doi: 10.1136/vr.134.26.681. [DOI] [PubMed] [Google Scholar]
  6. Cousins D. V., Williams S. N., Ross B. C., Ellis T. M. Use of a repetitive element isolated from Mycobacterium tuberculosis in hybridization studies with Mycobacterium bovis: a new tool for epidemiological studies of bovine tuberculosis. Vet Microbiol. 1993 Oct;37(1-2):1–17. doi: 10.1016/0378-1135(93)90178-a. [DOI] [PubMed] [Google Scholar]
  7. Groenen P. M., Bunschoten A. E., van Soolingen D., van Embden J. D. Nature of DNA polymorphism in the direct repeat cluster of Mycobacterium tuberculosis; application for strain differentiation by a novel typing method. Mol Microbiol. 1993 Dec;10(5):1057–1065. doi: 10.1111/j.1365-2958.1993.tb00976.x. [DOI] [PubMed] [Google Scholar]
  8. Hermans P. W., van Soolingen D., Bik E. M., de Haas P. E., Dale J. W., van Embden J. D. Insertion element IS987 from Mycobacterium bovis BCG is located in a hot-spot integration region for insertion elements in Mycobacterium tuberculosis complex strains. Infect Immun. 1991 Aug;59(8):2695–2705. doi: 10.1128/iai.59.8.2695-2705.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Hermans P. W., van Soolingen D., Dale J. W., Schuitema A. R., McAdam R. A., Catty D., van Embden J. D. Insertion element IS986 from Mycobacterium tuberculosis: a useful tool for diagnosis and epidemiology of tuberculosis. J Clin Microbiol. 1990 Sep;28(9):2051–2058. doi: 10.1128/jcm.28.9.2051-2058.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hermans P. W., van Soolingen D., van Embden J. D. Characterization of a major polymorphic tandem repeat in Mycobacterium tuberculosis and its potential use in the epidemiology of Mycobacterium kansasii and Mycobacterium gordonae. J Bacteriol. 1992 Jun;174(12):4157–4165. doi: 10.1128/jb.174.12.4157-4165.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Neill S. D., Pollock J. M., Bryson D. B., Hanna J. Pathogenesis of Mycobacterium bovis infection in cattle. Vet Microbiol. 1994 May;40(1-2):41–52. doi: 10.1016/0378-1135(94)90045-0. [DOI] [PubMed] [Google Scholar]
  12. O'Reilly L. M., Daborn C. J. The epidemiology of Mycobacterium bovis infections in animals and man: a review. Tuber Lung Dis. 1995 Aug;76 (Suppl 1):1–46. doi: 10.1016/0962-8479(95)90591-x. [DOI] [PubMed] [Google Scholar]
  13. Poulet S., Cole S. T. Characterization of the highly abundant polymorphic GC-rich-repetitive sequence (PGRS) present in Mycobacterium tuberculosis. Arch Microbiol. 1995 Feb;163(2):87–95. doi: 10.1007/BF00381781. [DOI] [PubMed] [Google Scholar]
  14. Poulet S., Cole S. T. Repeated DNA sequences in mycobacteria. Arch Microbiol. 1995 Feb;163(2):79–86. doi: 10.1007/BF00381780. [DOI] [PubMed] [Google Scholar]
  15. Ross B. C., Raios K., Jackson K., Dwyer B. Molecular cloning of a highly repeated DNA element from Mycobacterium tuberculosis and its use as an epidemiological tool. J Clin Microbiol. 1992 Apr;30(4):942–946. doi: 10.1128/jcm.30.4.942-946.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Skuce R. A., Brittain D., Hughes M. S., Beck L. A., Neill S. D. Genomic fingerprinting of Mycobacterium bovis from cattle by restriction fragment length polymorphism analysis. J Clin Microbiol. 1994 Oct;32(10):2387–2392. doi: 10.1128/jcm.32.10.2387-2392.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. van Embden J. D., Cave M. D., Crawford J. T., Dale J. W., Eisenach K. D., Gicquel B., Hermans P., Martin C., McAdam R., Shinnick T. M. Strain identification of Mycobacterium tuberculosis by DNA fingerprinting: recommendations for a standardized methodology. J Clin Microbiol. 1993 Feb;31(2):406–409. doi: 10.1128/jcm.31.2.406-409.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. van Soolingen D., Hermans P. W., de Haas P. E., Soll D. R., van Embden J. D. Occurrence and stability of insertion sequences in Mycobacterium tuberculosis complex strains: evaluation of an insertion sequence-dependent DNA polymorphism as a tool in the epidemiology of tuberculosis. J Clin Microbiol. 1991 Nov;29(11):2578–2586. doi: 10.1128/jcm.29.11.2578-2586.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. van Soolingen D., Hermans P. W., de Haas P. E., van Embden J. D. Insertion element IS1081-associated restriction fragment length polymorphisms in Mycobacterium tuberculosis complex species: a reliable tool for recognizing Mycobacterium bovis BCG. J Clin Microbiol. 1992 Jul;30(7):1772–1777. doi: 10.1128/jcm.30.7.1772-1777.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. van Soolingen D., de Haas P. E., Haagsma J., Eger T., Hermans P. W., Ritacco V., Alito A., van Embden J. D. Use of various genetic markers in differentiation of Mycobacterium bovis strains from animals and humans and for studying epidemiology of bovine tuberculosis. J Clin Microbiol. 1994 Oct;32(10):2425–2433. doi: 10.1128/jcm.32.10.2425-2433.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]

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