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Biochemistry (Moscow)

, Volume 69, Issue 5, pp 536–541 | Cite as

Comparative Analysis of Different Typing Methods for Helicobacter pylori Clinical Isolates

  • V. M. GovorunEmail author
  • P. G. Lokhov
  • S. A. Moshkovskii
  • K. T. Momynaliev
  • O. V. Selesnyova
  • L. V. Kudryavtseva
  • M. V. Serebryakova
  • O. V. Tikhonova
  • E. I. Goufman
  • A. I. Archakov
Article

Abstract

The goal of the present work was to compare different techniques of molecular typing using as an example clinical isolates of Helicobacter pylori obtained from patients in different regions of Russia. DNA-macroarray genome scanning using individual genes was employed to set up our basic classification of isolates that did or did not contain pathogenicity islands. In parallel, DNA of the same isolates was used in the conventional random amplified polymorphic DNA (RAPD) PCR analysis, and the isolates were also genotyped (cagA, vacA, iceA, and babA status) and their proteomic maps were obtained by means of unidimensional SDS polyacrylamide gel electrophoresis (1D-SDS-PAGE) coupled with identification using peptide mass fingerprinting by MALDI-TOF mass spectrometry. A statistically significant correlation (coefficient of correlation r = 0.25, p = 0.005) was observed between the results of genome scanning and 1D-SDS-PAGE. No correlation was found between RAPD-PCR typing and genome scanning.

Helicobacter pylori molecular typing genome scanning DNA array RAPD analysis proteomic map 

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REFERENCES

  1. 1.
    El-Omar, E. M., Chow, W. H., and Rabkin, C. S. (2001) Gastroenterology, 121, 1002-1004.Google Scholar
  2. 2.
    Meining, A., Riedl, B., and Stolte, M. (2002) J. Clin. Pathol., 55, 770-773.Google Scholar
  3. 3.
    Poundeyr, R. E., and Ng, D. (1995) Aliment. Pharmacol. Ther., 9 (Suppl. 2), 33-39.Google Scholar
  4. 4.
    Salama, N., Guillemin, K., McDaniel, T. K., Sherlock, G., Tompkins, L., and Falkow, S. (2000) Proc. Natl. Acad. Sci. USA, 97, 14668-14673.Google Scholar
  5. 5.
    Israel, D. A., Salama, N., Krishna, U., Rieger, U. M., Atherton, J. C., Falkow, S., and Peek, R. M., Jr. (2001) Proc. Natl. Acad. Sci. USA, 98, 14625-14630.Google Scholar
  6. 6.
    Israel, D. A., Salama, N., Arnold, C. N., Moss, S. F., Ando, T., Wirth, H. P., Tham, K. T., Camorlinga, M., Blaser, M. J., Falkow, S., and Peek, R. M., Jr. (2001) J. Clin. Invest., 107, 611-620.Google Scholar
  7. 7.
    Nomura, A. M., Lee, J., Stemmermann, G. N., Nomura, R. Y., Perez-Perez, G. I., and Blaser, M. J. (2002) J. Infect. Dis., 186, 1138-1144.Google Scholar
  8. 8.
    Tomb, J. F., White, O., Kerlavage, A. R., Clayton, R. A., Sutton, G. G., Fleischmann, R. D., Ketchum, K. A., Klenk, H. P., Gill, S., Dougherty, B. A., Nelson, K., Quackenbush, J., Zhou, L., Kirkness, E. F., Peterson, S., Loftus, B., Richardson, D., Dodson, R., Khalak, H. G., Glodek, A., McKenney, K., Fitzegerald, L. M., Lee, N., Adams, M. D., Venter, J. C., et al. (1997) Nature, 388, 539-547.Google Scholar
  9. 9.
    Alm, R. A., Ling, L. S., Moir, D. T., King, B. L., Brown, E. D., Doig, P. C., Smith, D. R., Noonan, B., Guild, B. C., deJonge, B. L., Carmel, G., Tummino, P. J., Caruso, A., Uria-Nickelsen, M., Mills, D. M., Ives, C., Gibson, R., Merberg, D., Mills, S. D., Jiang, Q., Taylor, D. E., Vovis, G. F., and Trust, T. J. (1999) Nature, 397, 176-180.Google Scholar
  10. 10.
    On, S. L., and Holmes, B. (1992) J. Clin. Microbiol., 30, 746-749.Google Scholar
  11. 11.
    Cover, T. L., and Blaser, M. J. (1992) J. Biol. Chem., 267, 10570-10575.Google Scholar
  12. 12.
    Van Doorn, L. J., Figueiredo, C., Sanna, R., and Pena, S. (1998) J. Clin. Microbiol., 36, 2597-2603.Google Scholar
  13. 13.
    VanDoorn, L. J., Figueiredo, C., Sanna, R., Plaisier, A., Schneeberger, P., deBoer, W., and Qoit, W. (1998) Gastroenterology, 115, 58-66.Google Scholar
  14. 14.
    VanDoorn, L. J., Henskens, Y., Nouhan, N., and Verschuuren, A. (2000) J. Clin. Microbiol., 38, 13-17.Google Scholar
  15. 15.
    Akopyants, N., Bukanov, N. O., Westblom, T. U., Kresovich, S., and Berg, D. E. (1992) Nucleic Acids Res., 20, 5137-5142.Google Scholar
  16. 16.
    Van der Ende, A., Pan, Z. J., Bart, A., van der Hulst, R. W., Feller, M., Xiao, S. D., Tytgat, G. N., and Dankert, J. (1998) Infection Immunity, 66, 1822-1826.Google Scholar
  17. 17.
    Van de Peer, Y., and de Wachter, R. (1997) Comput. Appl. Biosci., 13, 227-230.Google Scholar
  18. 18.
    Petersohn, A., Brigulla, M., Haas, S., Hoheisel, J. D., Volker, U., and Hecker, M. (2001) J. Bacteriol., 183, 5613-5617.Google Scholar
  19. 19.
    Laemmli, U. K. (1970) Nature, 227, 680-685.Google Scholar
  20. 20.
    Goerg, A., and Weiss, W. (1999) in 2-D Proteome Analysis Protocols (Methods in Molecular Biology) (Link, A. J., ed.) Vol. 112, Humana Press, Totowa, New Jersey, pp. 235-244.Google Scholar
  21. 21.
    Neuhoff, V., Arold, N., Taube, D., and Ehrhardt, W. (1988) Electrophoresis, 9, 255-262.Google Scholar
  22. 22.
    Shevchenko, A., Wilm, M., Vorm, O., and Mann, M. (1996) Analyt. Chem., 68, 850-858.Google Scholar
  23. 23.
    Govorun, V. M., Moshkovskii, S. A., Tikhonova, O. V., Goufman, E. I., Serebryakova, M. V., Momynaliev, K. T., Lokhov, P. G., Khryapova, E. V., Kudryavtseva, L. V., Smirnova, O. V., Toropyguine, I. Yu., Maksimov, B. I., and Archakov, A. I. (2003) Biochemistry (Moscow), 68, 42-49.Google Scholar
  24. 24.
    Ward, J. (1963) J. Am. Stat. Assoc., 58, 236-244.Google Scholar
  25. 25.
    Sokal, R. R., and Rohlf, F. J. (1995) Biometry, 3rd Edn., W. H. Freeman and Company, New York.Google Scholar
  26. 26.
    Mantel, N. (1967) Cancer Res., 27, 209-220.Google Scholar
  27. 27.
    Jungblut, P. R., Bumann, D., Haas, G., Zimny-Arndt, U., Holland, P., Lamer, S., Siejak, F., Aebischer, A., and Meyer, T. F. (2000) Mol. Microbiol., 36, 710-725.Google Scholar

Copyright information

© MAIK “Nauka/Interperiodica” 2004

Authors and Affiliations

  • V. M. Govorun
    • 1
    • 2
    Email author
  • P. G. Lokhov
    • 1
  • S. A. Moshkovskii
    • 1
  • K. T. Momynaliev
    • 2
  • O. V. Selesnyova
    • 2
  • L. V. Kudryavtseva
    • 2
  • M. V. Serebryakova
    • 1
  • O. V. Tikhonova
    • 1
  • E. I. Goufman
    • 1
  • A. I. Archakov
    • 1
  1. 1.Orekhovich Institute of Biomedical ChemistryRussian Academy of Medical SciencesMoscowRussia
  2. 2.Institute of Physico-Chemical MedicineMinistry of Public Health of Russian FederationMoscowRussia

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