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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

Melissa May

NetBio - Vice President – Strategic Planning

Date: 22/10/2013

slide3

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

090413

requirements ande rapid dna analysis system i
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
slide5

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
ande system components and process flow
ANDE System Components and Process Flow

Collection Swab

BioChipSet™ Cassette

Fully Automated

Instrument

Automated

Data Analysis

ande system specifications
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.
dna processing complete
DNA Processing Complete

Processing Complete

str analysis how does it work
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

str amplification what are short tandem repeats strs

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
str analysis how does it work1
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

dna profiling is extraordinarily accurate
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

traditional dna forensic laboratory infrastructure
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

five lab processes in one portable device
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

ande netbio s rapid dna instrument
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

field forward testing to mil std 810f
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
process flow dna purification
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

process flow rapid multiplexed str amplification
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

process flow electrophoretic separation and detection
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

data analysis and dna profile generation
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.
ande testing
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
rapid dna analysis an enabling technology
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
slide33

830 Winter Street, Waltham, MA 02451

www.netbio.com

Melissa.May@netbio.com

781-981-8273