• We are sorry, but NCBI web applications do not support your browser and may not function properly. More information
Logo of jcmPermissionsJournals.ASM.orgJournalJCM ArticleJournal InfoAuthorsReviewers
J Clin Microbiol. Jul 1992; 30(7): 1829–1836.
PMCID: PMC265389

Use of restriction fragment length polymorphisms resolved by pulsed-field gel electrophoresis for subspecies identification of mycobacteria in the Mycobacterium avium complex and for isolation of DNA probes.


Mycobacterial strains from the Mycobacterium avium complex were compared with each other and with Mycobacterium phlei isolates by restriction endonuclease digestion of chromosomal DNA with SspI and analysis by pulsed-field gel electrophoresis. Characteristic profiles were observed for known typed strains, and five groups were identified. Primary bovine isolates identified as Mycobacterium paratuberculosis by classical methods were shown to fall into both the M. paratuberculosis- and M. avium-like groups. M. paratuberculosis 18 was in the latter category. Two Mycobacterium intracellulare strains of different Schaefer serotypes had different digestion profiles. In addition, this system was exploited for the preparation of DNA probes by the isolation, digestion, and subcloning of DNA fragments separated by pulsed-field gel electrophoresis. Probe JC12 hybridized only to M. avium complex strains, but not to M. phlei, showing characteristic hybridization profiles for each of the groups previously identified by pulsed-field gel electrophoresis. The approach taken in the study lends itself to the comparative analysis of members of the M. avium complex and to the isolation and characterization of DNA probes with specificity for these mycobacteria.

Full text

Full text is available as a scanned copy of the original print version. Get a printable copy (PDF file) of the complete article (2.0M), or click on a page image below to browse page by page. Links to PubMed are also available for Selected References.

Images in this article

Click on the image to see a larger version.

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Anz W, Lauterbach D, Meissner G, Willers I. Vergleich von Sensitin-Testen an Meerschweinchen mit Serotyp und Hühnervirulenz bei M. avium- und M. intracellulare-Stämmen. Zentralbl Bakteriol Orig. 1970;215(4):536–549. [PubMed]
  • Baess I. Deoxyribonucleic acid relatedness among species of slowly-growing mycobacteria. Acta Pathol Microbiol Scand B. 1979 Aug;87(4):221–226. [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]
  • Camphausen RT, Jones RL, Brennan PJ. Antigenic relationship between Mycobacterium paratuberculosis and Mycobacterium avium. Am J Vet Res. 1988 Aug;49(8):1307–1310. [PubMed]
  • Chiodini RJ. Biochemical characteristics of various strains of Mycobacterium paratuberculosis. Am J Vet Res. 1986 Jul;47(7):1442–1445. [PubMed]
  • Chiodini RJ. The genetic relationship between Mycobacterium paratuberculosis and the M. avium complex. Acta Leprol. 1989;7 (Suppl 1):249–251. [PubMed]
  • Chiodini RJ. Crohn's disease and the mycobacterioses: a review and comparison of two disease entities. Clin Microbiol Rev. 1989 Jan;2(1):90–117. [PMC free article] [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, Thayer WR, Coutu JA, Merkal RS. In vitro antimicrobial susceptibility of a Mycobacterium sp. isolated from patients with Crohn's disease. Antimicrob Agents Chemother. 1984 Dec;26(6):930–932. [PMC free article] [PubMed]
  • Collins DM, Gabric DM, De Lisle GW. Identification of a repetitive DNA sequence specific to Mycobacterium paratuberculosis. FEMS Microbiol Lett. 1989 Jul 15;51(1):175–178. [PubMed]
  • Collins DM, Gabric DM, de Lisle GW. Identification of two groups of Mycobacterium paratuberculosis strains by restriction endonuclease analysis and DNA hybridization. J Clin Microbiol. 1990 Jul;28(7):1591–1596. [PMC free article] [PubMed]
  • Collins FM. AIDS-related mycobacterial disease. Springer Semin Immunopathol. 1988;10(4):375–391. [PubMed]
  • Drake TA, Hindler JA, Berlin OG, Bruckner DA. Rapid identification of Mycobacterium avium complex in culture using DNA probes. J Clin Microbiol. 1987 Aug;25(8):1442–1445. [PMC free article] [PubMed]
  • Ellner PD, Kiehn TE, Cammarata R, Hosmer M. Rapid detection and identification of pathogenic mycobacteria by combining radiometric and nucleic acid probe methods. J Clin Microbiol. 1988 Jul;26(7):1349–1352. [PMC free article] [PubMed]
  • Feinberg AP, Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem. 1983 Jul 1;132(1):6–13. [PubMed]
  • Good RC. Opportunistic pathogens in the genus Mycobacterium. Annu Rev Microbiol. 1985;39:347–369. [PubMed]
  • Hampson SJ, Portaels F, Thompson J, Green EP, Moss MT, Hermon-Taylor J, McFadden JJ. DNA probes demonstrate a single highly conserved strain of Mycobacterium avium infecting AIDS patients. Lancet. 1989 Jan 14;1(8629):65–68. [PubMed]
  • Lévy-Frébault VV, Thorel MF, Varnerot A, Gicquel B. DNA polymorphism in Mycobacterium paratuberculosis, "wood pigeon mycobacteria," and related mycobacteria analyzed by field inversion gel electrophoresis. J Clin Microbiol. 1989 Dec;27(12):2823–2826. [PMC free article] [PubMed]
  • McFadden JJ, Butcher PD, Chiodini R, Hermon-Taylor J. Crohn's disease-isolated mycobacteria are identical to Mycobacterium paratuberculosis, as determined by DNA probes that distinguish between mycobacterial species. J Clin Microbiol. 1987 May;25(5):796–801. [PMC free article] [PubMed]
  • McFadden JJ, Butcher PD, Chiodini RJ, Hermon-Taylor J. Determination of genome size and DNA homology between an unclassified Mycobacterium species isolated from patients with Crohn's disease and other mycobacteria. J Gen Microbiol. 1987 Jan;133(1):211–214. [PubMed]
  • McFadden JJ, Butcher PD, Thompson J, Chiodini R, Hermon-Taylor J. The use of DNA probes identifying restriction-fragment-length polymorphisms to examine the Mycobacterium avium complex. Mol Microbiol. 1987 Nov;1(3):283–291. [PubMed]
  • Meissner G, Schröder KH, Amadio GE, Anz W, Chaparas S, Engel HW, Jenkins PA, Käppler W, Kleeberg HH, Kubala E, et al. A co-operative numerical analysis of nonscoto- and nonphotochromogenic slowly growing mycobacteria. J Gen Microbiol. 1974 Aug;83(2):207–235. [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]
  • Picken RN, Plotch SJ, Wang Z, Lin BC, Donegan JJ, Yang HL. DNA probes for mycobacteria. I. Isolation of DNA probes for the identification of Mycobacterium tuberculosis complex and for mycobacteria other than tuberculosis (MOTT). Mol Cell Probes. 1988 Jun;2(2):111–124. [PubMed]
  • Picken RN, Tsang AY, Yang HL. Speciation of organisms within the Mycobacterium avium-Mycobacterium intracellulare-Mycobacterium scrofulaceum (MAIS) complex based on restriction fragment length polymorphisms. Mol Cell Probes. 1988 Dec;2(4):289–304. [PubMed]
  • Roberts MC, McMillan C, Coyle MB. Whole chromosomal DNA probes for rapid identification of Mycobacterium tuberculosis and Mycobacterium avium complex. J Clin Microbiol. 1987 Jul;25(7):1239–1243. [PMC free article] [PubMed]
  • Runyon EH. Pathogenic mycobacteria. Bibl Tuberc. 1965;21:235–287. [PubMed]
  • Saito H, Tomioka H, Sato K, Tasaka H, Dawson DJ. Identification of various serovar strains of Mycobacterium avium complex by using DNA probes specific for Mycobacterium avium and Mycobacterium intracellulare. J Clin Microbiol. 1990 Aug;28(8):1694–1697. [PMC free article] [PubMed]
  • Saito H, Tomioka H, Sato K, Tasaka H, Tsukamura M, Kuze F, Asano K. Identification and partial characterization of Mycobacterium avium and Mycobacterium intracellulare by using DNA probes. J Clin Microbiol. 1989 May;27(5):994–997. [PMC free article] [PubMed]
  • Saxegaard F, Baess I. Relationship between Mycobacterium avium, Mycobacterium paratuberculosis and "wood pigeon mycobacteria". Determinations by DNA-DNA hybridization. APMIS. 1988 Jan;96(1):37–42. [PubMed]
  • Sherman I, Harrington N, Rothrock A, George H. Use of a cutoff range in identifying mycobacteria by the Gen-Probe Rapid Diagnostic System. J Clin Microbiol. 1989 Feb;27(2):241–244. [PMC free article] [PubMed]
  • Sugden EA, Samagh BS, Bundle DR, Duncan JR. Lipoarabinomannan and lipid-free arabinomannan antigens of Mycobacterium paratuberculosis. Infect Immun. 1987 Mar;55(3):762–770. [PMC free article] [PubMed]
  • Thorel MF. Review of the occurrence of mycobactin dependence among mycobacteria species. Ann Rech Vet. 1984;15(3):405–409. [PubMed]
  • Thoresen OF, Saxegaard F. Gen-Probe Rapid Diagnostic System for the Mycobacterium avium complex does not distinguish between Mycobacterium avium and Mycobacterium paratuberculosis. J Clin Microbiol. 1991 Mar;29(3):625–626. [PMC free article] [PubMed]
  • Wards BJ, Collins DM, de Lisle GW. Restriction endonuclease analysis of members of the Mycobacterium avium-M. intracellulare-M. scrofulaceum serocomplex. J Clin Microbiol. 1987 Dec;25(12):2309–2313. [PMC free article] [PubMed]
  • Whipple D, Kapke P, Vary C. Identification of restriction fragment length polymorphisms in DNA from Mycobacterium paratuberculosis. J Clin Microbiol. 1990 Nov;28(11):2561–2564. [PMC free article] [PubMed]
  • Whipple DL, Le Febvre RB, Andrews RE, Jr, Thiermann AB. Isolation and analysis of restriction endonuclease digestive patterns of chromosomal DNA from Mycobacterium paratuberculosis and other Mycobacterium species. J Clin Microbiol. 1987 Aug;25(8):1511–1515. [PMC free article] [PubMed]
  • Yokomizo Y, Yugi H, Merkal RS. A method for avoiding false-positive reactions in an enzyme-linked immunosorbent assay (ELISA) for the diagnosis of bovine paratuberculosis. Nihon Juigaku Zasshi. 1985 Feb;47(1):111–119. [PubMed]
  • Yoshimura HH, Graham DY, Estes MK, Merkal RS. Investigation of association of mycobacteria with inflammatory bowel disease by nucleic acid hybridization. J Clin Microbiol. 1987 Jan;25(1):45–51. [PMC free article] [PubMed]

Articles from Journal of Clinical Microbiology are provided here courtesy of American Society for Microbiology (ASM)


Cited by other articles in PMC

See all...


Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...