title: Validation of high throughput sequencing and microbial forensics applications.
abstract: High throughput sequencing (HTS) generates large amounts of high quality sequence data for microbial genomics. The value of HTS for microbial forensics is the speed at which evidence can be collected and the power to characterize microbial-related evidence to solve biocrimes and bioterrorist events. As HTS technologies continue to improve, they provide increasingly powerful sets of tools to support the entire field of microbial forensics. Accurate, credible results allow analysis and interpretation, significantly influencing the course and/or focus of an investigation, and can impact the response of the government to an attack having individual, political, economic or military consequences. Interpretation of the results of microbial forensic analyses relies on understanding the performance and limitations of HTS methods, including analytical processes, assays and data interpretation. The utility of HTS must be defined carefully within established operating conditions and tolerances. Validation is essential in the development and implementation of microbial forensics methods used for formulating investigative leads attribution. HTS strategies vary, requiring guiding principles for HTS system validation. Three initial aspects of HTS, irrespective of chemistry, instrumentation or software are: 1) sample preparation, 2) sequencing, and 3) data analysis. Criteria that should be considered for HTS validation for microbial forensics are presented here. Validation should be defined in terms of specific application and the criteria described here comprise a foundation for investigators to establish, validate and implement HTS as a tool in microbial forensics, enhancing public safety and national security.
authors.affiliation: Department of Molecular and Medical Genetics, Institute of Applied Genetics, University of North Texas Health Science Center, Fort Worth, Texas, USA ; Center of Excellence in Genomic Medicine Research (CEGMR), King Abdulaziz University, Jeddah, Saudi Arabia.
authors.forename: Bruce
authors.initials: B
authors.lastname: Budowle
authors.affiliation: Rutgers New Jersey Medical School, Center for Biodefense, Rutgers University, Newark, New Jersey, USA.
authors.forename: Nancy D
authors.initials: ND
authors.lastname: Connell
authors.affiliation: Department of Epidemiology, The General K. Kaczkowski Military Institute of Hygiene and Epidemiology, Warsaw, Poland.
authors.forename: Anna
authors.initials: A
authors.lastname: Bielecka-Oder
authors.affiliation: CosmosID®, 387 Technology Dr, College Park, MD, USA ; Maryland Pathogen Research Institute, University of Maryland, College Park, MD, USA ; University of Maryland Institute for Advanced Computer Studies, University of Maryland, College Park, MD, USA ; Bloomberg School of Public Health, Johns Hopkins University, Baltimore, MD, USA.
authors.forename: Rita R
authors.initials: RR
authors.lastname: Colwell
authors.affiliation: Bioforensics Assay Development and DiagnosticsSection, Science Technology and Core Services Division, National Microbiology Laboratory, Winnipeg, MB, Canada ; Department of Medical Microbiology, University of Manitoba, Winnipeg, Canada.
authors.forename: Cindi R
authors.initials: CR
authors.lastname: Corbett
authors.affiliation: National Institute for Microbial Forensics & Food and Agricultural Biosecurity, Oklahoma State University, Stillwater, OK, USA.
authors.forename: Jacqueline
authors.initials: J
authors.lastname: Fletcher
authors.affiliation: Division of CBRN Defence and Security, Swedish Defence Research Agency, Umeå, Sweden.
authors.forename: Mats
authors.initials: M
authors.lastname: Forsman
authors.affiliation: Signature Science, LLC, Austin, TX, USA.
authors.forename: Dana R
authors.initials: DR
authors.lastname: Kadavy
authors.affiliation: University Hospital for Infectious Diseases "Fran Mihaljevic" and Medical School University of Rijeka, Zagreb, Croatia.
authors.forename: Alemka
authors.initials: A
authors.lastname: Markotic
authors.affiliation: Division of Foodborne, Waterborne, and Environmental Diseases, National Center for Emerging and Zoonotic Infectious Diseases, Centers for Disease Control and Prevention, Atlanta, Georgia.
authors.forename: Stephen A
authors.initials: SA
authors.lastname: Morse
authors.affiliation: Virginia Tech, National Capital Region, Arlington, VA, USA.
authors.forename: Randall S
authors.initials: RS
authors.lastname: Murch
authors.affiliation: Department of Molecular and Medical Genetics, Institute of Applied Genetics, University of North Texas Health Science Center, Fort Worth, Texas, USA ; Department of Forensic Medicine, Hjelt Institute, University of Helsinki, Helsinki, Finland.
authors.forename: Antti
authors.initials: A
authors.lastname: Sajantila
authors.affiliation: Department of Molecular and Medical Genetics, Institute of Applied Genetics, University of North Texas Health Science Center, Fort Worth, Texas, USA.
authors.forename: Sarah E
authors.initials: SE
authors.lastname: Schmedes
authors.affiliation: Signature Science, LLC, Austin, TX, USA.
authors.forename: Krista L
authors.initials: KL
authors.lastname: Ternus
authors.affiliation: Public Health Sciences, Bioinformatics Core Director, University of Virginia School of Medicine, Charlottesville, VA, USA.
authors.forename: Stephen D
authors.initials: SD
authors.lastname: Turner
authors.affiliation: Signature Science, LLC, Austin, TX, USA.
authors.forename: Samuel
authors.initials: S
authors.lastname: Minot
keywords: Bioinformatics; High throughput sequencing; Library preparation; Microbial forensics; Sample preparation; Validation
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