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Melissa May

Melissa May. NetBio - Vice President – Strategic Planning. Date: 22/10/2013. Heading. Fully automated, Field forward Rapid DNA Typing for Military, Intelligence, and Law Enforcement Applications. 090413. Requirements: ANDE Rapid DNA Analysis System (I).

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Melissa May

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  1. Melissa May NetBio - Vice President – Strategic Planning Date: 22/10/2013

  2. Heading

  3. Fully automated, Field forward Rapid DNA Typing for Military, Intelligence, and Law Enforcement Applications 090413

  4. Requirements: ANDE Rapid DNA Analysis System (I) In 2009 NetBio was awarded the ANDE contract, a competitive R&D program funded by a consortium of 5 Federal agencies to develop a Rapid DNA Analysis System based on: • Ease-of-use.The Operator has no technical training. • No manual processing • No reagent loading—reagents should be incorporated into the single-use consumable • No instrument calibration or assembly • No data interpretation—automated allele calling and searching • Reagent Stability/Out-of-the-box functionality. The System will be placed in a field forward environment. • No temperature, humidity, or particulate control • No refrigeration • No tools or technicians to install/align system • Generator power

  5. Requirements: ANDE Rapid DNA Analysis System (II) • Ruggedization. The system must be hardened for transport to and operation in deployed environments. • Rapid. DNA analysis must provide results within a time period that allows decisions to be made BEFORE releasing a person of interest. • Data security. All aspects of system operation and data generation must be secure. • User and sample tracking • Encryption of all generated data and database • Flexibility. The system must provide a customizable foundation for field forward forensic DNA analysis. • Touch sample analysis • Kinship analysis • Multi-assay capability

  6. ANDE System Components and Process Flow Collection Swab BioChipSet™ Cassette Fully Automated Instrument Automated Data Analysis

  7. ANDE System Specifications • Sample Types: Fresh and dried buccal and blood swabs, purified DNA. • Number of Samples Per Run: 5 plus allelic ladder. • Analysis Time: DNA profile in 84 minutes. • Operator Responsibility: Insert samples and initiate run. No other manipulations are required. • Reagent Stability: Reagents are stable for 6 months at room temperature, no refrigeration required. • Data Analysis: Fully automatic, integrated expert system software. No analyst review required. • Sample Tracking: Integrated RFID scanner on board the instrument allows tracking of RFID enables sample swabs. • System Maintenance: Field serviceable, annual maintenance. • Data Security: Password protected access and data encryption.

  8. Operating the ANDE System: Collect Samples

  9. Enter Sample ID

  10. Scan a Sample with the Integrated RFID Reader

  11. Insert Sample and Press the Done Button

  12. Load the Sample into the BioChipSet

  13. Press the Done Button

  14. Insert the BioChipSet Cassette

  15. Initiating the Run

  16. DNA Processing Complete Processing Complete

  17. Female DNA Profile

  18. STR Analysis, How Does It Work? • Unknown: Evidence Sample: • Blood, Body Fluids • Cigarette Butt • Handled Objects (cup, pop can, water bottle, etc) • Murder Weapon • Known: Reference Sample: • Convicted Offenders • Arrestees • Suspects • Paternity DNA Purification DNA Purification DNA Quantitation DNA Quantitation STR Amplification CE Separation & Detection DNA Profile Generation Database Search/Match

  19. AATG ~ ~ ~ ~ 7 repeats (maternal) 8 repeats (paternal) STR Amplification: What Are Short Tandem Repeats (STRs)? • Human DNA is 99% identical from person to person. However, some regions of the genome vary, and one type of variable region is the Short Tandem Repeat (STR). • Short Tandem Repeats (STR) are repetitive sequences: • Tetranucleotide: AATG AATG AATG AATG

  20. STR Analysis, How Does It Work? • Unknown: Evidence Sample: • Blood, Body Fluids • Cigarette Butt • Cup • Murder Weapon • Known: Reference Sample: • Convicted Offenders • Arrestees • Suspects • Paternity STR Amplification DNA Purification DNA Purification DNA Quantitation DNA Quantitation STR Amplification CE Separation & Detection DNA Profile Generation Database Search/Match

  21. Female DNA Profile

  22. DNA Profiling is Extraordinarily Accurate • J. Bulter, http://www.cstl.nist.gov/biotech/strbase/fbicore.htm The probability of a random match for this STR profile in the U.S. population is less than 1 in 837 trillion

  23. Traditional DNA Forensic Laboratory Infrastructure • Laboratory Infrastructure • Capital Equipment • Lab Supplies and Reagents • Computers and software • Laboratory Technicians • Forensic Analysts Sample Prep Lab Amplification Lab Separation & Detection Lab U.S. Crime Lab $210 M, 351,000 sqft DNA Unit ~$56 M, 70,000 sqft, annual operational budget >$2 M Forensic Analysis

  24. Five Lab Processes in One Portable Device Rapid DNA Analysis for Human Identification is the automated generation of STR profiles (DNA fingerprints) by non-technical operators outside the laboratory. Human AnalystProfile MatchHours - Weeks

  25. ANDE: NetBio’s Rapid DNA Instrument • Instrument Weight: 50 kg (two person lift) • Instrument Size: 68 x 42 x 59cm • Ruggedized to MIL STD 810F for shock / vibration • No calibration or realignment required after transport GPS Window Touchscreen Monitor BioChipSet Access Door Folding Carry Handle RFID Reader

  26. Field Forward Testing to MIL STD 810F • The ANDE System has under gone extensive ruggedization and field testing • Shock • Bench Handling • Restrained Cargo Vibration • Fragility • Operability with Generator Power

  27. Process Flow: DNA Purification Insert Swab (x5) Cell Lysis DNA Purification • All BioChipSet process steps are controlled using pneumatic pressures. • Guanidinium/Silica filter based purification. • Self-normalizing to delivery consistent range of DNA concentration for amplification. Elute DNA

  28. Process Flow: Rapid Multiplexed STR Amplification Purified DNA Sample Reconstitute PCR Cake Transfer to PCR Chambers • PCR uses Promega PowerPlex® 16 System. • PCR reagents have been lyophilized for long-term stability at room temperature (>2 years). • Rapid amplification: 31 cycles, 19.5 min. Amplification

  29. Process Flow: Electrophoretic Separation and Detection Join PCR Product & Formamide Reconstitute 5 ILS Cake/1 AL Injection 5 Samples/1 AL Electrophoretic Separation Fluorescence Detection

  30. Data Analysis and DNA Profile Generation • Expert System software performs automated data processing & allele designation. • Three data output files are automatically generated for database searching and • No data review or analysis by a forensic analyst is required.

  31. ANDE Testing • U.S. Testing Partners: USACIL, FBI, and NIST have been performing laboratory testing for the past 12 months • U.S. Military/Intelligence Exercise Trident Specter: deployed in maritime operations • SOCOM: initial laboratory testing performed, currently deployed for field testing • Australia: Successfully deployed in field exercise Q3 2013 • DHS: Immigration pilot scheduled for Q4 2013

  32. Rapid DNA Analysis: An Enabling Technology Rapid DNA Analysis for Human Identification is the automated generation of STR profiles (DNA fingerprints) by non-technical operators outside the laboratory. Major applications to enhance societal safety include: • Military and Intelligence Operations • Database Enrollment • Counter Terrorism/Counter Narcotics • Maritime Operations • Enemy Combatant Identification • Casualty identification • Homeland Security • Immigration processing • Mass Disaster Victim Identification • Borders and ports protection • Passport/document fraud • Human trafficking prevention • Law Enforcement • Arrestee processing • Crime laboratory support

  33. 830 Winter Street, Waltham, MA 02451 www.netbio.com Melissa.May@netbio.com 781-981-8273

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