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Characterization of Bacillus strains and hoax agents by protein profiling using automated microfluidic capillary electrophoresis

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Abstract

Purpose

In recent times, but especially since 2001, bioterrorism has been of increasing concern. In addition to the use of biological agents, including Bacillus anthracis (anthrax), there have been numerous hoax white powder “scares.” It is imperative to rapidly and accurately identify any suspicious powder as hazardous or hoax. Classical methods for identification typically rely on time-consuming cultivation or highly specific molecular tests which are limited if the agent is unknown. Faster and field portable methods for analysis of suspicious powders are urgently required.

Methods

Potential hoax agents, including Bacillus species and household powders, were analyzed using automated microfluidic capillary electrophoresis to determine if protein profiling can distinguish between, and identify, samples.

Results

Distinctive protein profiles were produced for Bacillus species, with the presence and/or absence of certain bands, aiding identification. In particular B. anthracis Sterne strain contained a distinctive doublet band above 100 kDa which was not present in any other Bacillus species or hoax agents examined. The majority of powders produced distinctive banding that could enable the identification of the sample while simultaneously ruling out B. anthracis with a high degree of confidence.

Conclusions

Results show automated microfluidic capillary electrophoresis can rapidly and reproducibly characterize Bacillus species and hoax agents based on protein profiles without the need for culture. Results were reproducible and there was enhanced resolution and rapidity compared to traditional protein profiling methods. Results show this technique is amenable to field use at a bioterrorism incident, thereby providing essential information to investigators regarding containment and treatment strategies.

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References

  1. The United States Department of Justice. Amerithrax investigative summary. Washington: The United States Department of Justice; 2010.

    Google Scholar 

  2. Leask A, Delpech V, McAnulty J. Anthrax and other suspect powders: Initial responses to an outbreak of hoaxes and scares. NSW Public Health Bull. 2003;14:218–21.

    Article  Google Scholar 

  3. Ash C, Farrow JAE, Dorsch M, Stackebrandt E, Collins MD. Comparative analysis of Bacillus anthracis, Bacillus cereus, and related species on the basis of reverse transcriptase sequencing of 16S rRNA. Int J Syst Bacteriol. 1991;41:343–6.

    Article  CAS  PubMed  Google Scholar 

  4. Ash C, Farrow JAE, Wallbanks S, Collins MD. Phylogenetic heterogeneity of the genus Bacillus revealed by comparative analysis of small-subunit-ribosomal RNA sequences. Lett Appl Microbiol. 1991;13:202–6.

    Article  CAS  Google Scholar 

  5. Ellerbrok H, Natterman H, Ozel M, Beutin L, Appel B, Pauli G. Rapid and sensitive identification of pathogenic and apathogenic Bacillus anthracis by real-time PCR. FEMS Microbiol Lett. 2002;214:51–9.

    Article  CAS  PubMed  Google Scholar 

  6. Oh SY, Budzik JM, Garufi G, Schneewind O. Two capsular polysaccharides enable Bacillus cereus G9241 to cause anthrax-like disease. Mol Microbiol. 2011;80:455–70.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  7. Mallik S, Virdi JS. Whole cell protein profiling reiterate phylogenetic relationships among strains of Yersinia enterocolitica biovar 1A as discerned earlier by different genotyping methods. J Appl Microbiol. 2010;109:946–52.

    Article  CAS  PubMed  Google Scholar 

  8. Sanchez I, Sesena S, Palop L. Identification of lactic acid bacteria from spontaneous fermentation of ‘Almagro’ eggplants by SDS-PAGE whole cell protein fingerprinting. Int J Food Microbiol. 2003;82:181–9.

    Article  CAS  PubMed  Google Scholar 

  9. Ayyadurai S, Flaudrops C, Raoult D, Drancourt M. Rapid identification and typing of Yersinia pestis and other Yersinia species by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry. BMC Microbiol. 2010;10:285.

    Article  PubMed Central  PubMed  Google Scholar 

  10. Barreiro JR, Braga PAC, Ferreira CR, Kostrzewa M, Maier T, Wegemann B, Boettcher V, Eberlin MN, dos Santos MV. Nonculture-based identification of bacteria in milk by protein fingerprinting. Proteomics. 2012;12:2739–45.

    Article  CAS  PubMed  Google Scholar 

  11. Ferreira L, Sanchez-Juanes F, Gonzalez-Avila M, Cembrero-Fucinos D, Herrero-Hernandez A, Gonzalez-Buitrago JM, Munoz-Bellido JL. Direct identification of urinary tract pathogens from urine samples by matrix-assisted laser desorption ionization-time of flight mass spectrometry. J Clin Microbiol. 2010;48:2110–5.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  12. Hsieh SJ, Tseng CL, Lee YS, Kuo AJ, Sun CF, Lin YH, Chen JK. Highly efficient classification and identification of human pathogenic bacteria by MALDI-TOF MS. Mol Cell Proteomics. 2008;7:448–56.

    Article  CAS  PubMed  Google Scholar 

  13. van Veen SQ, Claas ECJ, Kuijper EJ. High-throughput identification of bacteria and yeast by matrix-assisted laser desorption ionization–time of flight mass spectrometry in conventional medical microbiology laboratories. J Clin Microbiol. 2010;48:900–7.

    Article  PubMed Central  PubMed  Google Scholar 

  14. Pizarro SA, Lane P, Lane TW, Cruz E, Haroldsen B, VanderNoot VA. Bacterial characterization using protein profiling in a microchip separations platform. Electrophoresis. 2007;28:4697–704.

    Article  CAS  PubMed  Google Scholar 

  15. Stachowiak JC, Shugard EE, Mosier BP, Renzi RF, Caton PF, Ferko SM, Van de Vreugde JL, Yee DD, Haroldsen BL, VanderNoot VA. Autonomous microfluidic sample preparation system for protein profile-based detection of aerosolized bacterial cells and spores. Anal Chem. 2007;79:5763–70.

    Article  CAS  PubMed  Google Scholar 

  16. Drobniewski FA. Bacillus cereus and related species. Clin Microbiol Rev. 1993;6:324–38.

    CAS  PubMed Central  PubMed  Google Scholar 

  17. Minucci A, Delibato E, Castagnola M, Concolino P, Ameglio F, Zuppi C. Identification of RFLP G6PD mutations by using microcapillary electrophoretic chips (Experion). J Sep Sci. 2008;31:2694–700.

    Article  CAS  PubMed  Google Scholar 

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Correspondence to Michelle E. Gahan.

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McLaughlin, J., Nelson, M., McNevin, D. et al. Characterization of Bacillus strains and hoax agents by protein profiling using automated microfluidic capillary electrophoresis. Forensic Sci Med Pathol 10, 380–389 (2014). https://doi.org/10.1007/s12024-014-9578-z

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  • DOI: https://doi.org/10.1007/s12024-014-9578-z

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