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Effect of Sterilants on Amplification and Detection of Target DNA from Bacillus cereus Spores
Authors:James M Robertson PhD  Douglas L Anders PhD  Francene Basalyga MD  Julie Millar MFS  Donia Palomo Slack MS  Robert Bever PhD
Institution:1. Counterterrorism and Forensic Science Research Unit, FBI Laboratory, Quantico, VA;2. Scientific Response Analysis Unit, FBI Laboratory, Quantico, VA;3. Bode Cellmark Forensics, Inc., Lorton, VA;4. RTI International, Research Triangle Park, NC
Abstract:To conceal criminal activity of a bioterrorist or agroterrorist, the site of pathogen generation is often treated with sterilants to kill the organisms and remove evidence. As dead organisms cannot be analyzed by culture, this study examined whether DNA from sterilant‐treated Bacillus cereus spores was viable for amplification. The spores were exposed to five common sterilants: bleach, Sterilox®, oxidizer foam (L‐Gel), a peroxyacid (Actril®), and formaldehyde vapor. The spores were inoculated on typical surfaces found in offices and laboratories to test for environmental effects. It was found that the surface influenced the efficiency of recovery of the organisms. The DNA isolated from the recovered spores was successfully detected using RT‐qPCR for all treatments except for formaldehyde, by amplifying the phosphatidylinositol phospholipase C and sphingomyelinase genes. The results demonstrated that evidence from sites treated with sterilants can still provide information on the uncultured organism, using DNA amplification.
Keywords:forensic science  DNA amplification     Bacillus cereus     spores  sterilants  bleach  Actril  formaldehyde  Sterilox  L‐Gel
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