Validation of high throughput sequencing and microbial forensics applications

Bruce Budowle, Nancy D. Connell, Anna Bielecka-Oder, Rita R. Colwell, Cindi R. Corbett, Jacqueline Fletcher, Mats Forsman, Dana R. Kadavy, Alemka Markotic, Stephen A. Morse, Randall S. Murch, Antti Sajantila, Sarah E. Schmedes, Krista L. Ternus, Stephen D. Turner, Samuel Minot

Research output: Contribution to journalReview articlepeer-review

41 Scopus citations


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.

Original languageEnglish (US)
Article number9
JournalInvestigative Genetics
Issue number1
StatePublished - Jul 30 2014
Externally publishedYes


  • Bioinformatics
  • High throughput sequencing
  • Library preparation
  • Microbial forensics
  • Sample preparation
  • Validation

ASJC Scopus subject areas

  • Pathology and Forensic Medicine
  • Molecular Biology
  • Genetics


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