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Applied and Environmental Microbiology, October 2009, p. 6157-6167, Vol. 75, No. 19
0099-2240/09/$08.00+0     doi:10.1128/AEM.00766-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Novel and Unique Diagnostic Biomarkers for Bacillus anthracis Infection{triangledown}

Sagit Sela-Abramovich,2,{dagger} Theodor Chitlaru,1,{dagger} Orit Gat,1 Haim Grosfeld,1 Ofer Cohen,1 and Avigdor Shafferman1*

Department of Biochemistry and Molecular Genetics, Israel Institute for Biological Research,1 Life Science Research Israel Ltd., Ness-Ziona 74100, Israel2

Received 5 April 2009/ Accepted 22 July 2009

A search for bacterium-specific biomarkers in peripheral blood following infection with Bacillus anthracis was carried out with rabbits, using a battery of specific antibodies generated by DNA vaccination against 10 preselected highly immunogenic bacterial antigens which were identified previously by a genomic/proteomic/serologic screen of the B. anthracis secretome. Detection of infection biomarkers in the circulation of infected rabbits could be achieved only after removal of highly abundant serum proteins by chromatography using a random-ligand affinity column. Besides the toxin component protective antigen, the following three secreted proteins were detected in the circulation of infected animals: the chaperone and protease HtrA (BA3660), an NlpC/P60 endopeptidase (BA1952), and a protein of unknown function harboring two SH3 (Src homology 3) domains (BA0796). The three proteins could be detected in plasma samples from infected animals exhibiting 103 to 105 CFU/ml blood and also in standard blood cultures at 3 to 6 h post-bacterial inoculation at a bacteremic level as low as 103 CFU/ml. Furthermore, the three biomarkers appear to be present only in the secretome of B. anthracis, not in those of the related pathogens B. thuringiensis and B. cereus. To the best of our knowledge, this is the first report of direct detection of B. anthracis-specific proteins, other than the toxin components, in the circulation of infected animals.

* Corresponding author. Mailing address: Israel Institute for Biological Research, P.O. Box 19, Ness-Ziona 74100, Israel. Phone: 972 8 9381595. Fax: 972 8 9401404. E-mail: avigdors{at}iibr.gov.il

{triangledown} Published ahead of print on 31 July 2009.

{dagger} S.S.-A. and T.C. contributed equally to this study.

Applied and Environmental Microbiology, October 2009, p. 6157-6167, Vol. 75, No. 19
0099-2240/09/$08.00+0     doi:10.1128/AEM.00766-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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