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Environmental Survey for Four Pathogenic Bacteria and Closely Related Species Using Phylogenetic and Functional Genes

NCJ Number
Journal of Forensic Sciences Volume: 51 Issue: 3 Dated: May 2006 Pages: 548-558
Cheryl R. Kuske Ph.D.; Susan M. Barns Ph.D.; Christy C. Grow B.S.; Lori Merrill M.S.; John Dunbar Ph.D.
Date Published
May 2006
11 pages
Since bacterial species similar to pathogens (agents that cause disease) may have a similar DNA sequence, this study determined the natural presence of four pathogenic bacteria and their closely related species for a large collection of soil and aerosol samples.
Close relatives of Bacillus anthracis (Bacillus cereus group species) were found in 37 percent of the soils and 25 percent of the aerosol samples. The Bacillus anthracis protective antigen (pag) gene or a close homolog was found in 16 of these samples. For the other three pathogen groups--Clostridium perfringens, Francisella tularensis, and Yersinia pestis--the frequency of detection was much lower, and none of the samples were positive with both the phylogenetic and virulence gene primer sets. Soil samples (approximately 10-50 cubic cm) were collected from 0-10 cm of surface soils or shoreline sediments of ponds/lakes. Aerosol filters were obtained from U.S. Environmental Protection Agency State air quality monitoring agencies. Primers for the amplification of 16S rRNA genes from each of the four pathogens and their close relatives were designed for the study or were modified from primers described in the literature. This report also describes the design and testing of virulence gene primer sets, PCR amplification, the cloning and sequencing of PCR products, and sequence data analysis. 3 tables and 65 references