GIFT Generic Integrated Forensic Toolbox for CBRN incidents

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GIFT Generic Integrated Forensic Toolbox for CBRN incidents Presentation vid Aktörsgemensamt CBRNE-möte Stockholm, 25 jan 2018 Forskningsledare, Martin Nygren FOI Funded under the seventh Framework Program of the EC

Disposition Hur började det hela? GIFT vilken typ av projekt var det? Projektstruktur/arbetspaket/deltagare Resultat från arbetspaket 2 och 5

EU-utlysning EU-utlysning: FP7-SEC 2013-1 Utlysningsdatum: 10 juli 2012 Inlämningsdatum: 22 nov 2012 kl. 17:00 Ämnesområde: European toolbox, focusing on procedures, practices and guidelines for CBRN forensic aspects Projektavslut: Avslutande möte den 16 okt 2017 (171130) Budget: 3,5 miljoner Euro

Konsortium och projektidé Möte på NFI, Holland, 11 okt 2012 Partners många ville vara med (myndigheter, företag) Projektidén var Forensik med frågeställningarna Vad har hänt? Vad orsakade det? Spreds några farliga ämnen ut? Vem gjorde det? Är vi redo att undersöka brottsplatsen? Skulle vi ha kunnat förhindra händelsen? Terrorister eller kriminella? Industriolycka?

Mål och syfte med GIFT The project aims to make advancements in forensic research by providing procedures practicalities innovative tools and equipment and best practices in doing forensic research on CBRN contaminated incident scenes methods for traditional forensic research on contaminated evidence and profiling the CBRN agents released Finally, a forensic toolbox for the forensic investigators acting on the incident scene and the forensic researchers in the laboratories.

GIFT - Struktur Huvudområden som projektansökan skulle innehålla Scenarier Brottsplatsundersökning Traditionell forensiska analyser på kontaminerat material CBRN forensik Toolbox Utvärdering, övningar och utbildning Legala, etiska och sociala aspekter Resultatspridning

GIFT - Struktur WP 1 Management Lead: NFI WP2 Threat assessment and tool box design WP3 Crime Scene Investigation WP4 Tradidional forensics on cont. mtrl. WP5 CBRNe forensic investigation WP6 Integration of forensic toolbox WP7 Validation, exercise & education WP8 Legal, ethical and social aspects WP9 Dessimination & exploatation Lead: TNO Lead: RIVM Lead: NFI Lead: FOI Lead:Tyndall Lead: RIVM Lead: Eticas Lead: CBRNw Task 2:1 Scenarios Task 2.2 State of the art Task 3.2 Forensic sampling (FOI/SKL) Task 3.3 Chain of custody Task 4.1-4.5 Decontamination of material Total budget 7 116 000 Projekttid 3 år Rapporter Milstolpar 87 st. 26 st. FOI budget 903 000 Deltagare (partners): Länder Myndigheter Små företag varav Kriminaltekniska laboratorier Möten (WP 5) 21 st. 9 st. 12 st. 9 st. 3 st. 15 st.

Arbetspaket 2 (WP 2) Threat assessment and toolbox design Task 2.1 CBRN scenarios Task 2.2 State of the Art for Forensic Investigation Task 2.3 Future situation for the short- and long term Task 2.4 Re-evaluation of the existing Gap Analysis Task 2.5 Input for CBBRN forensic Toolbox Design FOI deltog i två delaktiviteter som genererade 3 st. rapporter 1. CBRN scenarios for the forensic community innehåller 7 C, 8 B, 6 R och 2 N scenarier (Restricted) 2. Overview of relevant historic incidents 3. Integrated scenario report (Restricted)

Syftet med arbetspaket 5 (WP 5) Syftet med arbetspaket fem vara att utveckla laboratoriemetoder för CBRN profilering och använda dessa signaturer för att bättre kunna identifiera källan till CBRN-ämnena och möjligen också binda brottet till en förövare

Struktur WP 5

Leveranser WP 5 Deliverable D 5.1 D 5.2.1 D 5.2.2R D 5.3 D 5.4 D 5.5 D 5.6 D 5.7 D 5.8 D 5.9.1R D 5.9.2 Title Establishment of analytical tools for Chemical Threat Agents analysis Establishment of analytical tools for Toxin analysis Toxin Analysis, Restricted Annex Sampling strategy for microbial forensic investigation Evaluation of methods for source attribution of a microbial pathogen using simulated data of genomic signatures Final report on Chemical Forensic Analysis of Chemical Threat Agents Final report on Chemical Forensic Analysis of Toxins Sample preparation procedures for molecular analysis of bio agents in a forensic context Final report on use of genomic signatures for attribution purposes Report on current capabilities available at European laboratories Report on Nuclear Forensic of nuclear materials and signatures of radiological materials

Subtask 5.1.2 Toxins Aim Development of generic analytical tools for toxin, i.e. toxic biomolecules from living organisms (e.g. plants, fungi and moulds). Objectives General screening method for toxins. Attribution profiling of toxin samples. Perform a study using Amanita mushrooms as model. A complex set of attribution markers collected: Analysis of genetic markers Toxin and metabolite markers Stable isotope ratio- and trace metal analyses Attribution profiling of ama- and phallotoxins in food samples. Vit flugsvamp Lömsk flugsvamp Fool s mushroom

Subtask 5.1.2 Toxins Results A high resolution LC-MS screening method has been developed for screening of model toxins, spiked to different sample matrices. Attribution profiling of toxin samples with genetic and chemical analytical tools Toxic Amanita mushroom species used as model Species specific toxin profiles Geopositioning of source organisms.

Subtask 5.1.2 Toxins Highlights Rapid toxin analysis techniques based on LC-HRMS is good general tool for screening of samples for content of non-protein toxins Forensic profiling of toxin samples is useful for retrospective determination of source species Chemical analysis of chemical markers from the source organism and trace metal elements is a powerful tool for determination of the geographic origin of toxin source species

Subtask 5.2.1 DNA preparation in Bio agents Aim The aim of this subtask is to develop a tool which comprises a sampling and successive DNA-extraction step, that yields a non-infectious DNA/RNA-extract out of different samples which can be found at a crime scene or other relevant sites. The obtained non-infectious DNA/RNA extract has to meet the criteria which are needed for detection, identification or strain typing of B-agents. Tasks - Sampling strategy for microbial forensic investigation - Sampling preparation procedures for molecular analysis of bio agents in a forensic context - Requirements on sample preparation for different techniques

Subtask 5.2.1 DNA preparation in Bio agents Protocol to obtain a non-infectious DNA-extract (NFI) Tested at NFI: Lactobacillus casei (non-pathogenic bacteria) Protocol to obtain a non-infectious DNA extract starting with : 1. DNA-extraction (hand held bead beater) followed by 2. Centrifugal filtration 3. In this study a new application for HDP as a biological disinfectant was proposed. The tested HDP showed bactericidal activity and PCR amplifiable DNA was obtained after a DNA purification step. Methods 1 and 2 were efficient in killing or removal of L. casei cells in solutions up to 10 8 CFU ml -1 and yielded PCR amplifiable DNA.

Subtask 5.2.1 DNA preparation in Bio agents Proof of principle in during B-exercise in the Netherlands Amplifiable non-infectious DNA was obtained, the protocol works A: Control, 50 µl Yakult B: obtained DNA -extract(s)

Subtask 5.2.2 Identification of genomic signatures for improved attribution of bio agents Objective To provide statistical methods for calculating likelihood based evidence, which can be used in legal proceedings of source attribution of a bacterial pathogen used in an illegitimate context Lab A Lab B NGS How to? Evidence value +4 0-4 Scale used in general forensics to communicate results to court proceedings

Subtask 5.2.2 Identification of genomic signatures for improved attribution of bio agents Highlighted result The experiment with biological amplification of low frequency mutations revealed strong selection for mutations encoding cell surface structures important when differentiate natural/deliberate outbreak Mutation frequency Time