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. 1996 Dec;34(12):3044–3048. doi: 10.1128/jcm.34.12.3044-3048.1996

Evaluation of method for secondary DNA typing of Mycobacterium tuberculosis with pTBN12 in epidemiologic study of tuberculosis.

Z Yang 1, F Chaves 1, P F Barnes 1, W J Burman 1, J Koehler 1, K D Eisenach 1, J H Bates 1, M D Cave 1
PMCID: PMC229457  PMID: 8940446

Abstract

Secondary fingerprinting of Mycobacterium tuberculosis DNA with a probe containing the polymorphic GC-rich repetitive sequence present in pTBN12 has been found to have greater discriminating power than does fingerprinting with the insertion sequence IS6110 for strains carrying few copies of IS6110. To validate the use of pTBN12 fingerprinting in the molecular epidemiology of tuberculosis, M. tuberculosis isolates from 67 patients in five states in the United States and in Spain were fingerprinted with both IS6110 and pTBN12. Epidemiologic links among the 67 patients were evaluated by patient interview and/or review of medical records. The 67 isolates had 5 IS6110 fingerprint patterns with two to five copies of IS6110 and 18 pTBN12 patterns, of which 10 were shared by more than 1 isolate. Epidemiologic links are consistently found among patients whose isolates had identical pTBN12 patterns, whereas no links were found among patients whose isolates had unique pTBN12 patterns. This suggests that pTBN12 fingerprinting is a useful tool to identify epidemiologically linked tuberculosis patients whose isolates have identical IS6110 fingerprints containing fewer than six fragments.

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Selected References

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  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. Chaves F., Yang Z., el Hajj H., Alonso M., Burman W. J., Eisenach K. D., Dronda F., Bates J. H., Cave M. D. Usefulness of the secondary probe pTBN12 in DNA fingerprinting of Mycobacterium tuberculosis. J Clin Microbiol. 1996 May;34(5):1118–1123. doi: 10.1128/jcm.34.5.1118-1123.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Chevrel-Dellagi D., Abderrahman A., Haltiti R., Koubaji H., Gicquel B., Dellagi K. Large-scale DNA fingerprinting of Mycobacterium tuberculosis strains as a tool for epidemiological studies of tuberculosis. J Clin Microbiol. 1993 Sep;31(9):2446–2450. doi: 10.1128/jcm.31.9.2446-2450.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. 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]
  6. Das S., Paramasivan C. N., Lowrie D. B., Prabhakar R., Narayanan P. R. IS6110 restriction fragment length polymorphism typing of clinical isolates of Mycobacterium tuberculosis from patients with pulmonary tuberculosis in Madras, south India. Tuber Lung Dis. 1995 Dec;76(6):550–554. doi: 10.1016/0962-8479(95)90533-2. [DOI] [PubMed] [Google Scholar]
  7. Doran T. J., Hodgson A. L., Davies J. K., Radford A. J. Characterisation of a novel repetitive DNA sequence from Mycobacterium bovis. FEMS Microbiol Lett. 1992 Sep 15;75(2-3):179–185. doi: 10.1016/0378-1097(92)90400-i. [DOI] [PubMed] [Google Scholar]
  8. 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]
  9. 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]
  10. Genewein A., Telenti A., Bernasconi C., Mordasini C., Weiss S., Maurer A. M., Rieder H. L., Schopfer K., Bodmer T. Molecular approach to identifying route of transmission of tuberculosis in the community. Lancet. 1993 Oct 2;342(8875):841–844. doi: 10.1016/0140-6736(93)92698-s. [DOI] [PubMed] [Google Scholar]
  11. Godfrey-Faussett P., Mortimer P. R., Jenkins P. A., Stoker N. G. Evidence of transmission of tuberculosis by DNA fingerprinting. BMJ. 1992 Jul 25;305(6847):221–223. doi: 10.1136/bmj.305.6847.221. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. 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]
  13. 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]
  14. 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]
  15. 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]
  16. 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]
  17. 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]
  18. McAdam R. A., Hermans P. W., van Soolingen D., Zainuddin Z. F., Catty D., van Embden J. D., Dale J. W. Characterization of a Mycobacterium tuberculosis insertion sequence belonging to the IS3 family. Mol Microbiol. 1990 Sep;4(9):1607–1613. doi: 10.1111/j.1365-2958.1990.tb02073.x. [DOI] [PubMed] [Google Scholar]
  19. 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]
  20. Otal I., Martín C., Vincent-Lévy-Frebault V., Thierry D., Gicquel B. Restriction fragment length polymorphism analysis using IS6110 as an epidemiological marker in tuberculosis. J Clin Microbiol. 1991 Jun;29(6):1252–1254. doi: 10.1128/jcm.29.6.1252-1254.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. 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]
  22. 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]
  23. 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]
  24. Yang Z. H., de Haas P. E., Wachmann C. H., van Soolingen D., van Embden J. D., Andersen A. B. Molecular epidemiology of tuberculosis in Denmark in 1992. J Clin Microbiol. 1995 Aug;33(8):2077–2081. doi: 10.1128/jcm.33.8.2077-2081.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Yang Z. H., de Haas P. E., van Soolingen D., van Embden J. D., Andersen A. B. Restriction fragment length polymorphism Mycobacterium tuberculosis strains isolated from Greenland during 1992: evidence of tuberculosis transmission between Greenland and Denmark. J Clin Microbiol. 1994 Dec;32(12):3018–3025. doi: 10.1128/jcm.32.12.3018-3025.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. 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]
  27. Zainuddin Z. F., Dale J. W. Polymorphic repetitive DNA sequences in Mycobacterium tuberculosis detected with a gene probe from a Mycobacterium fortuitum plasmid. J Gen Microbiol. 1989 Sep;135(9):2347–2355. doi: 10.1099/00221287-135-9-2347. [DOI] [PubMed] [Google Scholar]
  28. 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]
  29. van Embden J. D., van Soolingen D., Small P. M., Hermans P. W. Genetic markers for the epidemiology of tuberculosis. Res Microbiol. 1992 May;143(4):385–391. doi: 10.1016/0923-2508(92)90051-o. [DOI] [PubMed] [Google Scholar]
  30. 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]
  31. 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]

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