1. NetBio - Vice President – Strategic Planning
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
For first bullet: As discussed in the previous talk, the system must be ruggedized

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

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
Repeat this 5x

13. Press the Done Button

14. Insert the BioChipSet Cassette
Point of scanning is sample tracking within the chip

15. Initiating the Run

16. DNA 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. 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
7 repeats (maternal)
8 repeats (paternal)
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
DNA Purification
DNA Quantitation
STR Amplification
CE Separation & Detection
DNA Profile Generation
Database
Search/Match
STR Amplification

21. Female DNA Profile

22. DNA Profiling is Extraordinarily Accurate
Locus
Allele
Value
1 in
Combined
D5S818 11
0.2533 13
0.2152
9.17 9
VWA 17
0.2815
8.87 81
FGA 25
0.1854 26
0.2185
12.35
1005
D8S1179 14
0.1656
16.29
16,364
D21S11 28
0.1589 31
0.2782
11.31
185,073
D18S51
0.1374
0.1391
26.18
4,845,217
D3S1358 15
0.3841 16
0.1407
9.25
44,818,259
D13S317 10
0.3394
0.0480
30.69
1.38 x 109
D7S820
0.1772
31.85
4.38 x 1010
D16S539
0.1126
0.3212
13.80
6.05 x 1011
THO1 6
0.2318
0.2814
18.62
1.13 x 1013
TPOX 8
0.5348
3.50
3.94 x 1013
CSF1PO
0.2169 12
21.28
8.37 x 1014
Regions of chromosomes that are looked at have no phenotypical information-except for gender.
This table looks complicated, but makes the point that as you interogate more regions of the genome the probability of a random match
J. Bulter,
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
Capital Equipment
Lab Supplies and Reagents
Computers and software
Laboratory Technicians
Forensic Analysts
Sample Prep Lab
Amplification Lab
Separation & Detection Lab
Most Foresnic Analysts have a masters degree and require 6 months of training, continuing education in a forensic lab before reaching full staff.
Washing dishes, and autoclaving for sterilization,
U.S. Crime Lab $210 M, 351,000 sq ft
DNA Unit ~$56 M, 70,000 sq ft, 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.
Rapid DNA Analysis for Human Identification is the automated generation of STR profiles (DNA fingerprints) by non-technical operators outside the laboratory.
Human Analyst Profile Match Hours - Weeks

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

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
Elute DNA
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.

28. Process Flow: Rapid Multiplexed STR Amplification
Purified DNA Sample
Reconstitute PCR Cake
Transfer to PCR Chambers
Amplification
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.

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
Highlight both kinship applications and linking people to crimes

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