Skip to main content
PMC home page
Journal of Clinical Microbiology logoLink to Journal of Clinical Microbiology
. 1996 May;34(5):1118–1123. doi: 10.1128/jcm.34.5.1118-1123.1996

Usefulness of the secondary probe pTBN12 in DNA fingerprinting of Mycobacterium tuberculosis.

F Chaves 1, Z Yang 1, H el Hajj 1, M Alonso 1, W J Burman 1, K D Eisenach 1, F Dronda 1, J H Bates 1, M D Cave 1
PMCID: PMC228966  PMID: 8727887

Abstract

A comparison was made between DNA fingerprints of Mycobacterium tuberculosis produced with the insertion sequence IS6110 and those produced with the polymorphic GC-rich repetitive sequence contained in the plasmid pTBN12. A total of 302 M. tuberculosis isolates from the prison system in Madrid, Spain, and the Denver Public Health Department (Denver, Colo.) were analyzed with the two probes. Both probes identified the same isolates in the same clusters when the fingerprints had six or more copies of IS6110. Analysis of isolates with unique IS6110 fingerprints demonstrated that they were unique with pTBN12. The pTBN12 probe had greater discriminating power in isolates having five or fewer copies of IS6110. Forty-seven isolates from Denver having fewer than five copies of IS6110 which were grouped in 11 clusters with identical fingerprint patterns were subdivided into 35 different patterns by pTBN12. Isolates with IS6110 fingerprints with more than six copies of IS6110 that differed from one another by only one or two hybridizing bands were analyzed with pTBN12. Most of these sets of isolates demonstrated identical patterns with pTBN12. However, some exceptions were observed, suggesting that those having nearly identical IS6110 patterns should not necessarily be included in the same cluster. Since IS6110 provides more polymorphism in the fingerprint, it is most useful in identifying isolates with unique fingerprint patterns and those in clusters in which the isolates contain six or more copies of the insertion. However, it is necessary to employ a secondary probe, such as pTBN12, to discriminate isolates with five or fewer copies of IS6110 and those with similar but not identical IS6110 patterns.

Full Text

The Full Text of this article is available as a PDF (714.3 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Alland D., Kalkut G. E., Moss A. R., McAdam R. A., Hahn J. A., Bosworth W., Drucker E., Bloom B. R. Transmission of tuberculosis in New York City. An analysis by DNA fingerprinting and conventional epidemiologic methods. N Engl J Med. 1994 Jun 16;330(24):1710–1716. doi: 10.1056/NEJM199406163302403. [DOI] [PubMed] [Google Scholar]
  2. Cave M. D., Eisenach K. D., McDermott P. F., Bates J. H., Crawford J. T. IS6110: conservation of sequence in the Mycobacterium tuberculosis complex and its utilization in DNA fingerprinting. Mol Cell Probes. 1991 Feb;5(1):73–80. doi: 10.1016/0890-8508(91)90040-q. [DOI] [PubMed] [Google Scholar]
  3. Cave M. D., Eisenach K. D., Templeton G., Salfinger M., Mazurek G., Bates J. H., Crawford J. T. Stability of DNA fingerprint pattern produced with IS6110 in strains of Mycobacterium tuberculosis. J Clin Microbiol. 1994 Jan;32(1):262–266. doi: 10.1128/jcm.32.1.262-266.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Dwyer B., Jackson K., Raios K., Sievers A., Wilshire E., Ross B. DNA restriction fragment analysis to define an extended cluster of tuberculosis in homeless men and their associates. J Infect Dis. 1993 Feb;167(2):490–494. doi: 10.1093/infdis/167.2.490. [DOI] [PubMed] [Google Scholar]
  5. Edlin B. R., Tokars J. I., Grieco M. H., Crawford J. T., Williams J., Sordillo E. M., Ong K. R., Kilburn J. O., Dooley S. W., Castro K. G. An outbreak of multidrug-resistant tuberculosis among hospitalized patients with the acquired immunodeficiency syndrome. N Engl J Med. 1992 Jun 4;326(23):1514–1521. doi: 10.1056/NEJM199206043262302. [DOI] [PubMed] [Google Scholar]
  6. Eisenach K. D., Crawford J. T., Bates J. H. Repetitive DNA sequences as probes for Mycobacterium tuberculosis. J Clin Microbiol. 1988 Nov;26(11):2240–2245. doi: 10.1128/jcm.26.11.2240-2245.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Feinberg A. P., Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem. 1983 Jul 1;132(1):6–13. doi: 10.1016/0003-2697(83)90418-9. [DOI] [PubMed] [Google Scholar]
  8. Hermans P. W., Messadi F., Guebrexabher H., van Soolingen D., de Haas P. E., Heersma H., de Neeling H., Ayoub A., Portaels F., Frommel D. Analysis of the population structure of Mycobacterium tuberculosis in Ethiopia, Tunisia, and The Netherlands: usefulness of DNA typing for global tuberculosis epidemiology. J Infect Dis. 1995 Jun;171(6):1504–1513. doi: 10.1093/infdis/171.6.1504. [DOI] [PubMed] [Google Scholar]
  9. 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]
  10. Mazurek G. H., Cave M. D., Eisenach K. D., Wallace R. J., Jr, Bates J. H., Crawford J. T. Chromosomal DNA fingerprint patterns produced with IS6110 as strain-specific markers for epidemiologic study of tuberculosis. J Clin Microbiol. 1991 Sep;29(9):2030–2033. doi: 10.1128/jcm.29.9.2030-2033.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Murray M. G., Thompson W. F. Rapid isolation of high molecular weight plant DNA. Nucleic Acids Res. 1980 Oct 10;8(19):4321–4325. doi: 10.1093/nar/8.19.4321. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Rastogi N., Ross B. C., Dwyer B., Goh K. S., Clavel-Sérès S., Jeantils V., Cruaud P. Emergence during unsuccessful chemotherapy of multiple drug resistance in a strain of Mycobacterium tuberculosis. Eur J Clin Microbiol Infect Dis. 1992 Oct;11(10):901–907. doi: 10.1007/BF01962370. [DOI] [PubMed] [Google Scholar]
  13. 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]
  14. Small P. M., Hopewell P. C., Singh S. P., Paz A., Parsonnet J., Ruston D. C., Schecter G. F., Daley C. L., Schoolnik G. K. The epidemiology of tuberculosis in San Francisco. A population-based study using conventional and molecular methods. N Engl J Med. 1994 Jun 16;330(24):1703–1709. doi: 10.1056/NEJM199406163302402. [DOI] [PubMed] [Google Scholar]
  15. Small P. M., Shafer R. W., Hopewell P. C., Singh S. P., Murphy M. J., Desmond E., Sierra M. F., Schoolnik G. K. Exogenous reinfection with multidrug-resistant Mycobacterium tuberculosis in patients with advanced HIV infection. N Engl J Med. 1993 Apr 22;328(16):1137–1144. doi: 10.1056/NEJM199304223281601. [DOI] [PubMed] [Google Scholar]
  16. Tabet S. R., Goldbaum G. M., Hooton T. M., Eisenach K. D., Cave M. D., Nolan C. M. Restriction fragment length polymorphism analysis detecting a community-based tuberculosis outbreak among persons infected with human immunodeficiency virus. J Infect Dis. 1994 Jan;169(1):189–192. doi: 10.1093/infdis/169.1.189. [DOI] [PubMed] [Google Scholar]
  17. Thierry D., Cave M. D., Eisenach K. D., Crawford J. T., Bates J. H., Gicquel B., Guesdon J. L. IS6110, an IS-like element of Mycobacterium tuberculosis complex. Nucleic Acids Res. 1990 Jan 11;18(1):188–188. doi: 10.1093/nar/18.1.188. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Yuen L. K., Ross B. C., Jackson K. M., Dwyer B. Characterization of Mycobacterium tuberculosis strains from Vietnamese patients by Southern blot hybridization. J Clin Microbiol. 1993 Jun;31(6):1615–1618. doi: 10.1128/jcm.31.6.1615-1618.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. 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]
  20. 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]
  21. van Soolingen D., de Haas P. E., Hermans P. W., Groenen P. M., van Embden J. D. Comparison of various repetitive DNA elements as genetic markers for strain differentiation and epidemiology of Mycobacterium tuberculosis. J Clin Microbiol. 1993 Aug;31(8):1987–1995. doi: 10.1128/jcm.31.8.1987-1995.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES