Abstract
This report describes the development of in-house real-time PCR assays using minor groove binding probes for simultaneous detection of the Bacillus anthracis pag and cap genes, the Francisella tularensis 23 KDa gene, as well as the Yersinia pestis pla gene. The sensitivities of these assays were at least 1 fg, except for the assay targeting the Bacillus anthracis cap gene, which showed a sensitivity of 10 fg when total DNA was used as a template in a serial dilution. The clinical value of the Bacillus anthracis- and Francisella tularensis-specific assays was demonstrated by successful amplification of DNA from cases of cow anthrax and hare tularemia, respectively. No cross-reactivity between these species-specific assays or with 39 other bacterial species was noted. These assays may provide a rapid tool for the simultaneous detection and identification of the three category A bacterial species listed as biological threats by the Centers for Disease Control and Prevention.
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Acknowledgements
This study was supported by a grant from the Finnish Scientific Advisory Board for Defence. We would like to thank Professor M. Vaara (HUSLAB, Helsinki, Finland) and Dr. M. Forsman (FOI, Swedish Defence Research Agency, Umeå, Sweden) for providing clinical cultures of F. tularensis and the F. tularensis LVS strain, respectively. Dr. M. Salminen (National Public Health Institute, Helsinki) is acknowledged for providing the laboratory facilities used for these studies. We also thank H. Hemmilä for excellent technical assistance and Dr. C. Heckman, as well as Ms. E. Nevalainen for editorial assistance. These experiments comply with the current laws of Finland.
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Skottman, T., Piiparinen, H., Hyytiäinen, H. et al. Simultaneous real-time PCR detection of Bacillus anthracis, Francisella tularensis and Yersinia pestis . Eur J Clin Microbiol Infect Dis 26, 207–211 (2007). https://doi.org/10.1007/s10096-007-0262-z
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DOI: https://doi.org/10.1007/s10096-007-0262-z