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.

Next generation sequencing (NGS): a golden tool in forensic toolkit

“…Umożliwia opracowywanie charakterystyki próbek na wysokim poziomie swoistości, z uwzględnieniem identyfikacji danego mikroorganizmu na różnych poziomach -gatunku, szczepu, izolatu, a nawet naczynia hodowlanego, z którego pochodzi prób-ka. Mikroorganizmy odgrywają także istotną rolę w procesach rozkładu [25]. Pechal i wsp.…”

“…Rozwojowi mikrobiologii sądowej sprzyjają także technologie sekwencjonowania wysokiej przepustowości (HTS), dostarczając tej dziedzinie skutecznych narzędzi [25]. Ostatnie badania wyjaśniono, że różne systemy bazujące na technologii NGS umożliwiają identyfikację śladów biologicznych oraz podejrzanych [3].…”

“…Recent studies elucidated that different systems of NGS technology has the abilities for identification of biological traces and suspect [3]. Furthermore, it able to detect microorganisms in low abundance and differentiate them from others by genomic signatures [25]. It is also highly sensitive in tracing genomic differences among isolates so it is a better biomarker than PCR probes, which target limited information [28,29].…”

“…Attribution is a sample characterization with high specificity, including identification of a microorganism at different levels such as species, strain, isolate or even the culture vessel from which the sample originated. Moreover, microbes play an important role in decomposition [25]. Pechal et al (2013) elucidated that bacterial communities in skin and mouth followed a consistent change during decomposition [26].…”

“…As high-throughput sequencing (HTS) technologies continue in introducing powerful tools in microbial forensics [25]. Recent studies elucidated that different systems of NGS technology has the abilities for identification of biological traces and suspect [3].…”