title: Whole-Genome Sequencing in Microbial Forensic Analysis of Gamma-Irradiated Microbial Materials.
abstract: Effective microbial forensic analysis of materials used in a potential biological attack requires robust methods of morphological and genetic characterization of the attack materials in order to enable the attribution of the materials to potential sources and to exclude other potential sources. The genetic homogeneity and potential intersample variability of many of the category A to C bioterrorism agents offer a particular challenge to the generation of attributive signatures, potentially requiring whole-genome or proteomic approaches to be utilized. Currently, irradiation of mail is standard practice at several government facilities judged to be at particularly high risk. Thus, initial forensic signatures would need to be recovered from inactivated (nonviable) material. In the study described in this report, we determined the effects of high-dose gamma irradiation on forensic markers of bacterial biothreat agent surrogate organisms with a particular emphasis on the suitability of genomic DNA (gDNA) recovered from such sources as a template for whole-genome analysis. While irradiation of spores and vegetative cells affected the retention of Gram and spore stains and sheared gDNA into small fragments, we found that irradiated material could be utilized to generate accurate whole-genome sequence data on the Illumina and Roche 454 sequencing platforms.
authors.affiliation: BioSciences Division, Research & Technology Directorate, Edgewood Chemical Biological Center, Aberdeen Proving Ground, Aberdeen, Maryland, USA.
authors.forename: Stacey M
authors.initials: SM
authors.lastname: Broomall
authors.affiliation: BioSciences Division, Research & Technology Directorate, Edgewood Chemical Biological Center, Aberdeen Proving Ground, Aberdeen, Maryland, USA Excet, Inc., Springfield, Virginia, USA.
authors.forename: Mohamed
authors.initials: M
authors.lastname: Ait Ichou
authors.affiliation: BioSciences Division, Research & Technology Directorate, Edgewood Chemical Biological Center, Aberdeen Proving Ground, Aberdeen, Maryland, USA Excet, Inc., Springfield, Virginia, USA.
authors.forename: Michael D
authors.initials: MD
authors.lastname: Krepps
authors.affiliation: BioSciences Division, Research & Technology Directorate, Edgewood Chemical Biological Center, Aberdeen Proving Ground, Aberdeen, Maryland, USA.
authors.forename: Lauren A
authors.initials: LA
authors.lastname: Johnsky
authors.affiliation: BioSciences Division, Research & Technology Directorate, Edgewood Chemical Biological Center, Aberdeen Proving Ground, Aberdeen, Maryland, USA.
authors.forename: Mark A
authors.initials: MA
authors.lastname: Karavis
authors.affiliation: BioSciences Division, Research & Technology Directorate, Edgewood Chemical Biological Center, Aberdeen Proving Ground, Aberdeen, Maryland, USA Excet, Inc., Springfield, Virginia, USA.
authors.forename: Kyle S
authors.initials: KS
authors.lastname: Hubbard
authors.affiliation: BioSciences Division, Research & Technology Directorate, Edgewood Chemical Biological Center, Aberdeen Proving Ground, Aberdeen, Maryland, USA Leidos Inc., Aberdeen Proving Ground, Aberdeen, Maryland, USA.
authors.forename: Joseph M
authors.initials: JM
authors.lastname: Insalaco
authors.affiliation: BioSciences Division, Research & Technology Directorate, Edgewood Chemical Biological Center, Aberdeen Proving Ground, Aberdeen, Maryland, USA.
authors.forename: Janet L
authors.initials: JL
authors.lastname: Betters
authors.affiliation: BioSciences Division, Research & Technology Directorate, Edgewood Chemical Biological Center, Aberdeen Proving Ground, Aberdeen, Maryland, USA.
authors.forename: Brady W
authors.initials: BW
authors.lastname: Redmond
authors.affiliation: BioSciences Division, Research & Technology Directorate, Edgewood Chemical Biological Center, Aberdeen Proving Ground, Aberdeen, Maryland, USA Leidos Inc., Aberdeen Proving Ground, Aberdeen, Maryland, USA.
authors.forename: Bryan A
authors.initials: BA
authors.lastname: Rivers
authors.affiliation: BioSciences Division, Research & Technology Directorate, Edgewood Chemical Biological Center, Aberdeen Proving Ground, Aberdeen, Maryland, USA Optimetrics Inc., Abingdon, Maryland, USA.
authors.forename: Alvin T
authors.initials: AT
authors.lastname: Liem
authors.affiliation: BioSciences Division, Research & Technology Directorate, Edgewood Chemical Biological Center, Aberdeen Proving Ground, Aberdeen, Maryland, USA Optimetrics Inc., Abingdon, Maryland, USA.
authors.forename: Jessica M
authors.initials: JM
authors.lastname: Hill
authors.affiliation: BioSciences Division, Research & Technology Directorate, Edgewood Chemical Biological Center, Aberdeen Proving Ground, Aberdeen, Maryland, USA Optimetrics Inc., Abingdon, Maryland, USA.
authors.forename: Edward T
authors.initials: ET
authors.lastname: Fochler
authors.affiliation: BioSciences Division, Research & Technology Directorate, Edgewood Chemical Biological Center, Aberdeen Proving Ground, Aberdeen, Maryland, USA Optimetrics Inc., Abingdon, Maryland, USA.
authors.forename: Pierce A
authors.initials: PA
authors.lastname: Roth
authors.affiliation: BioSciences Division, Research & Technology Directorate, Edgewood Chemical Biological Center, Aberdeen Proving Ground, Aberdeen, Maryland, USA.
authors.forename: C Nicole
authors.initials: CN
authors.lastname: Rosenzweig
authors.affiliation: BioSciences Division, Research & Technology Directorate, Edgewood Chemical Biological Center, Aberdeen Proving Ground, Aberdeen, Maryland, USA.
authors.forename: Evan W
authors.initials: EW
authors.lastname: Skowronski
authors.affiliation: BioSciences Division, Research & Technology Directorate, Edgewood Chemical Biological Center, Aberdeen Proving Ground, Aberdeen, Maryland, USA henry.s.gibbons.civ@mail.mil.
authors.forename: Henry S
authors.initials: HS
authors.lastname: Gibbons
mesh_terms: D001419:Bacteria / Q000235:genetics / Q000254:growth & development / Q000528:radiation effects*; D054045:Biological Warfare Agents; D044707:Forensic Sciences; D005720:Gamma Rays; D016680:Genome, Bacterial / Q000528:radiation effects*; D017422:Sequence Analysis, DNA
chemical_list: D054045:Biological Warfare Agents