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A method for differentiation of Bacillus anthracis strains and phylogenetically related species based on determination of the structural differences between chromosomal genes for biosynthesis of flagellin and methionine

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Abstract

Nucleotide sequence analysis of several genes responsible for definitive properties of the anthrax pathogen—motility and penicillinase activity—determined a chromosomal locus promising for interspecies differentiation. We demonstrated that the fliC gene encoding flagellin synthesis contains an extended region distinguishing B. anthracis strains from the majority of nonpathogenic and opportunistic bacilli. A novel method for anthrax pathogen indication and identification based on determination of the differences in the fliC and hom2 chromosomal genes structure has been proposed. A total of 60 strains of different Bacillus spp. (B. anthracis, B. cereus, B. thuringiensis, B. mycoides, B. megaterium, B. subtilis, etc.) were tested using two chromosomal DNA targets. The algorithm developed in this work permits detection of the pathogenic microorganism and reliably differentiation of it from other Bacillus spp. representatives. The introduction of primers complementary to specific sequences of pXO1 and pXO2 plasmids into multiplex PCR makes it possible to obtain additional information on the proposed virulence of the isolate.

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References

  1. Eremenko, E.I., Ryazanova, A.G., Buravtseva, N.P., Tsygankova, O.I., Aksenova, L.Yu., Antyuganov, S.N., et al., Analysis of anthrax incidence in 2012, the forecast for 2013, Probl. Osobo Opasn. Infekts., 2013, vol. 1, pp. 18–20.

    Google Scholar 

  2. Tourasse, N., Helgason, E., Okstad, O., Hegna, I., and Kolsto, A., The Bacillus cereus group: Novel aspects of population structure and genome dynamics, J. Appl. Microbiol., 2006, vol. 101, pp. 579–593.

    Article  CAS  PubMed  Google Scholar 

  3. Avashia, S., Riggins, W., Lindley, C., Hoffmaster, A., Drumgoole, R., Nekomoto, T., et al., Fatal pneumonia among metalworkers due to inhalation exposure to Bacillus cereus containing Bacillus anthracis toxin genes, Clin. Infect. Dis., 2007, vol. 44, pp. 414–416.

    Article  PubMed  Google Scholar 

  4. Hoffmaster, A., Ravel, J., Rasko, D., Chapman, G., Chute, M., Marston, C., et al., Identification of anthrax toxin genes in a Bacillus cereus associated with an illness resembling inhalation anthrax, Proc. Natl. Acad. Sci. USA, 2004, vol. 101, pp. 8449–8454.

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  5. Klee, S., Brzuszkiewicz, E., Nattermann, H., Brüggemann, H., Dupke, S., Wollherr, A., et al., The genome of a Bacillus isolate causing anthrax in chimpanzees combines chromosomal properties of B. cereus with B. anthracis virulence plasmids, PLoS One, 2010, vol. 5, no. 7, p. e10986.

    Article  PubMed Central  PubMed  Google Scholar 

  6. Easterday, R., Van Ert, M., Simonson, T., Wagner, D., Kenefic, L., Allender, C., and Keim, P., Use of single nucleotide polymorphisms in the plcR gene for specific identification of Bacillus anthracis, J. Clin. Microbiol., 2005, vol. 43, no. 4, pp. 1995–1997.

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  7. Hurtle, W., Bode, E., Kulesh, D., Kaplan, R., Garrison, J., Bridge, D., et al., Detection of the Bacillus anthracis gyrA gene by using a minor groove binder probe, J. Clin. Microbiol., 2004, vol. 42, pp. 179–185.

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  8. Patra, G., Sylvestre, P., Ramisse, V., Thérasse, J., and Guesdon, J., Isolation of a specific chromosomic DNA sequence of Bacillus anthracis and its possible use in diagnosis, FEMS Immunol. Med. Microbiol., 1996, vol. 15, pp. 223–231.

    Article  CAS  PubMed  Google Scholar 

  9. Qi, Y., Patra, G., Liang, X., Williams, L., Rose, S., Redkar, R., and DelVecchio, V., Utilization of the rpoB gene as a specific chromosomal marker for real-time PCR detection of Bacillus anthracis, Appl. Environ. Microbiol., 2001, vol. 67, no. 8, pp. 3720–3727.

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  10. Patra, G., Vaissaire, J., Weber-Levy, M., Le Doujet, C., and Mock, M., Molecular characterization of Bacillus strains involved in outbreaks of anthrax in France in 1997, J. Clin. Microbiol., 1998, vol. 36, pp. 3412–3414.

    PubMed Central  CAS  PubMed  Google Scholar 

  11. Zasada, A., Gierczynski, R., Raddadi, N., Daffonchio, D., and Jagielsi, M., Some Bacillus thuringiensis strains share rpoB nucleotide polymorphisms also present in Bacillus anthracis, J. Clin. Microbiol., 2006, vol. 44, pp. 1606–1607.

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  12. Irenge, L., Durant, J., Tomaso, H., Pilo, P., Olsen, J., Ramisse, V., et al., Development and validation of realtime quantitative PCR assay for rapid identification of Bacillus anthracis in environmental samples, Appl. Microbiol. Biotechnol., 2010, vol. 88, pp. 1179–1192.

    Article  CAS  PubMed  Google Scholar 

  13. Wielinga, P., Hamidjaja, R., Ågren, J., Knutsson, R., Segerman, B., Fricker, M., et al., A multiplex real-time PCR for identifying and differentiating B. anthracis virulent types, Int. J. Food Microbiol., 2011, vol. 145, pp. 135–144.

    Article  Google Scholar 

  14. Ahmod, N., Gupta, R., and Shah, H., Identification of a Bacillus anthracis specific indel in the yeaC gene and development of a rapid pyrosequencing assay for distinguishing B. anthracis from the B. cereus group, J. Microbiol. Methods, 2011, vol. 87, pp. 278–285.

    Article  CAS  PubMed  Google Scholar 

  15. Mikshis, N.I., Zhivova, Yu.N., Novikova, L.V., Sharapova, N.A., Popov, Yu.A., and Kutyrev, V.V., Identification of differences in structure of methionine biosynthesis genes in Bacillus anthracis strains and phylogenetically related species of bacilli, Mol. Genet., Microbiol. Virol., 2012, vol. 27, no. 2, p.p. 69–73.

    Article  Google Scholar 

  16. Ramisse, V., Patra, G., Garrigue, H., Guesdon, J., and Mock, M., Identification and characterization of Bacillus anthracis by multiplex PCR analysis of sequences on plasmids pXO1 and pXO2 and chromosomal DNA, FEMS Microbiol. Lett., 1996, vol. 145, pp. 9–16.

    Article  CAS  PubMed  Google Scholar 

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Correspondence to N. I. Mikshis.

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Original Russian Text © N.I. Mikshis, T.N. Kashtanova, V.V. Kutyrev, 2015, published in Molekulyarnaya Genetika, Mikrobiologiya i Virusologiya, 2015, No. 4, pp. 22–26.

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Mikshis, N.I., Kashtanova, T.N. & Kutyrev, V.V. A method for differentiation of Bacillus anthracis strains and phylogenetically related species based on determination of the structural differences between chromosomal genes for biosynthesis of flagellin and methionine. Mol. Genet. Microbiol. Virol. 30, 181–185 (2015). https://doi.org/10.3103/S0891416815040060

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  • DOI: https://doi.org/10.3103/S0891416815040060

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