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J Clin Microbiol. Aug 1987; 25(8): 1511–1515.
PMCID: PMC269259

Isolation and analysis of restriction endonuclease digestive patterns of chromosomal DNA from Mycobacterium paratuberculosis and other Mycobacterium species.


A relatively rapid and efficient method for the extraction of chromosomal DNA from Mycobacterium paratuberculosis and other mycobacteria was developed. Approximately 25 to 50 micrograms of DNA could be extracted from 100 mg (wet weight) of cells, which was sufficient to perform several restriction endonuclease analyses from a single preparation. The DNA from five Mycobacterium species, including four strains of M. paratuberculosis and four strains of M. avium, was analyzed by this method. Digestion with the restriction endonucleases BstEII and PstI yielded the most definitive restriction patterns. For some strains, the restriction endonuclease analysis results were in agreement with the current identification of these organisms. The two strains of M. avium serotype 2 had identical fragment patterns. Similarly, the two strains of M. avium complex serotype 6 had identical fragment patterns. The three mycobactin-dependent M. paratuberculosis strains were very similar, whereas the mycobactin-independent M. paratuberculosis strain was more similar to the M. avium serotype 2 strains. Although many more cultures would need to be evaluated to determine correct groupings, the results of this study demonstrated the potential of restriction enzyme analysis for the differentiation of slowly growing mycobacteria.

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Selected References

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  • Baess I. Isolation and purification of deoxyribonucleic acid from mycobacteria. Acta Pathol Microbiol Scand B Microbiol Immunol. 1974 Dec;82(6):780–784. [PubMed]
  • Baess I. Deoxyribonucleic acid relationships between different serovars of Mycobacterium avium, Mycobacterium intracellulare and Mycobacterium scrofulaceum. Acta Pathol Microbiol Immunol Scand B. 1983 Jun;91(3):201–203. [PubMed]
  • Chiodini RJ. Biochemical characteristics of various strains of Mycobacterium paratuberculosis. Am J Vet Res. 1986 Jul;47(7):1442–1445. [PubMed]
  • Chiodini RJ, Van Kruiningen HJ, Merkal RS. Ruminant paratuberculosis (Johne's disease): the current status and future prospects. Cornell Vet. 1984 Jul;74(3):218–262. [PubMed]
  • Chiodini RJ, Van Kruiningen HJ, Merkal RS, Thayer WR, Jr, Coutu JA. Characteristics of an unclassified Mycobacterium species isolated from patients with Crohn's disease. J Clin Microbiol. 1984 Nov;20(5):966–971. [PMC free article] [PubMed]
  • Collins DM, De Lisle GW. DNA restriction endonuclease analysis of Mycobacterium tuberculosis and Mycobacterium bovis BCG. J Gen Microbiol. 1984 Apr;130(4):1019–1021. [PubMed]
  • Collins DM, de Lisle GW. Restriction endonuclease analysis of various strains of Mycobacterium paratuberculosis isolated from cattle. Am J Vet Res. 1986 Oct;47(10):2226–2229. [PubMed]
  • Garcia MJ, Tabarés E. Separation of Mycobacterium gadium from other rapidly growing mycobacteria on the basis of DNA homology and restriction endonuclease analysis. J Gen Microbiol. 1986 Aug;132(8):2265–2269. [PubMed]
  • Le Febvre RB, Foley JW, Thiermann AB. Rapid and simplified protocol for isolation and characterization of leptospiral chromosomal DNA for taxonomy and diagnosis. J Clin Microbiol. 1985 Oct;22(4):606–608. [PMC free article] [PubMed]
  • Marshall RB, Wilton BE, Robinson AJ. Identification of Leptospira serovars by restriction-endonuclease analysis. J Med Microbiol. 1981 Feb;14(1):163–166. [PubMed]
  • Matthews PR, McDiarmid A. The production in bovine calves of a disease resembling paratuberculosis with a Mycobacterium sp isolated from a woodpigeon (Columba palumbus L). Vet Rec. 1979 Mar 31;104(13):286–286. [PubMed]
  • Matthews PR, McDiarmid A, Collins P, Brown A. The dependence of some strains of Mycobacterium avium on mycobactin for initial and subsequent growth. J Med Microbiol. 1978 Feb;11(1):53–57. [PubMed]
  • O'Hara MJ, Collins DM, de Lisle GW. Restriction endonuclease analysis of Brucella ovis and other Brucella species. Vet Microbiol. 1985 Aug;10(5):425–429. [PubMed]
  • Patel R, Kvach JT, Mounts P. Isolation and restriction endonuclease analysis of mycobacterial DNA. J Gen Microbiol. 1986 Feb;132(2):541–551. [PubMed]
  • Shoemaker SA, Fisher JH, Jones WD, Jr, Scoggin CH. Restriction fragment analysis of chromosomal DNA defines different strains of Mycobacterium tuberculosis. Am Rev Respir Dis. 1986 Aug;134(2):210–213. [PubMed]
  • Shoemaker SA, Fisher JH, Scoggin CH. Techniques of DNA hybridization detect small numbers of mycobacteria with no cross-hybridization with non-mycobacterial respiratory organisms. Am Rev Respir Dis. 1985 May;131(5):760–763. [PubMed]
  • Thiermann AB, Handsaker AL, Moseley SL, Kingscote B. New method for classification of leptospiral isolates belonging to serogroup pomona by restriction endonuclease analysis: serovar kennewicki. J Clin Microbiol. 1985 Apr;21(4):585–587. [PMC free article] [PubMed]
  • Wayne LG, Gross WM. Isolation of deoxyribonucleic acid from mycobacteria. J Bacteriol. 1968 Apr;95(4):1481–1482. [PMC free article] [PubMed]

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