Transcript Slide 1

DNA Extraction
for Telemedicine Security
Tofy Mussivand, FRSC
Professor of Surgery & Engineering
Chair, Medical Devices Centre, University of Ottawa
Director, Cardiovascular Devices Division, University of Ottawa Heart Institute
Working Hypothesis
A portable, rapid, device based system for
DNA extraction from invasive and non-invasive
(e.g. fingerprints) samples can be developed for
wide-ranging applications in both medical
and non-medical fields.
Dr. T. Mussivand, FRSC
Medical Devices Centre at the
University of Ottawa Heart Institute
Required Functions/Steps
For Device Based DNA System
1) Cell sample collection
(from fingerprint or other sources)
2) Lysis of cell membrane
3) Extraction of (nuclear) DNA
4) Purification of DNA
5) Restriction enzyme operation
(Cleavage of DNA strings into recognition sequence)
6) PCR (Polymerase chain reaction)
Replication and amplification of DNA
7) DNA detection
Dr. T. Mussivand, FRSC
and identification
Medical Devices Centre at the
University of Ottawa Heart Institute
DNA Testing Delays
News Headlines
• April 2005 - Washington State University researchers
uncover huge unsolved crime DNA testing backlog
(Washington State University News Service)
• May 2006 - Delays in obtaining forensic evidence
adversely affecting justice system
(Auditor General, Australian Government)
• May 2007 - DNA labs facing huge backlog:
Canadians and Americans failing to meet target
processing times…
(The Scientist)
• Feb 2008 - Lansing serial killing suspect's trial
delayed for DNA analysis
(WSBT News)
Dr. T. Mussivand, FRSC
Medical Devices Centre at the
University of Ottawa Heart Institute
Major Delays
• Royal Canadian Mounted Police (RCMP):
– Average turnaround time: 114 days
– Target turnaround time: 30 days
• Federal Bureau of Investigation (FBI):
– Average turnaround time: 1 year
– Target turnaround time: 60 days.
Source: The Scientist. May 21, 2007
Dr. T. Mussivand, FRSC
Medical Devices Centre at the
University of Ottawa Heart Institute
1st Question: Can DNA be obtained from
non-invasive sources (fingerprints) ?
Skin debris deposited on a plastic
surface after finger applied
Developed technique “Sample
Release” to lift the skin debris
Same surface following application
of the “Sample Release” technique
Light Microscope X 20 Magnification
Dr. T. Mussivand, FRSC
Medical Devices Centre at the
University of Ottawa Heart Institute
Next Question: Can sufficient DNA
be extracted from skin debris ?
60
50
DNA
40
Extracted
From Single 30
Fingerprints
20
(ng)
10
0
Working Threshold (10 ng)
1
2
3
4
5
6
7
8
9
10 11 12 13 14 15 16
Individual Test Subjects (volunteers)
Dr. T. Mussivand, FRSC
Medical Devices Centre at the
University of Ottawa Heart Institute
Next Question: Can this DNA sampling
and extraction be performed rapidly ?
16 hrs.
16
Processing
Time for DNA
Sampling &
Extraction
(hours)
13 hrs.
12
8
4
15 min.
0
Developed
Extraction
Technique
45 min.
Chelex 100
Extraction
Phenol-chloroform
Extraction
Salting out
Extraction
Existing Extraction Methods
Dr. T. Mussivand, FRSC
Medical Devices Centre at the
University of Ottawa Heart Institute
DNA Sampling & Extraction
Process Validation
• Process results
validated by an
independent 3rd
party:
– National DNA Data
Bank operated by the
Royal Canadian
Mounted Police
(RCMP)
Dr. T. Mussivand, FRSC
Medical Devices Centre at the
University of Ottawa Heart Institute
More Validation
Dr. T. Mussivand, FRSC
Medical Devices Centre at the
University of Ottawa Heart Institute
Current Focus
Integration of
Developed Techniques
into a Portable Device
Based System
Dr. T. Mussivand, FRSC
Medical Devices Centre at the
University of Ottawa Heart Institute
First Challenge: Microfluidics based
Cell Membrane Lysis to Extract DNA
A 10 sec. video
clip taken with a
20X microscope
Control Cell
The membrane of the cell
exposed to this newly developed
technique is lysed within
~ 6 seconds,
while the control cell is not lysed
Cell exposed to
newly developed
technique
Dr. T. Mussivand, FRSC
Medical Devices Centre at the
University of Ottawa Heart Institute
Next Steps: Cell Membrane Lysis using a new
microchip developed by the Medical Devices Centre



MUMPs Technology or Multi
User MEMS processes
MEMS = micro-electromechanical systems
MetalMUMPs = 20 micron
thick nickel electroplating
construction process
cm
Dr. T. Mussivand, FRSC
Medical Devices Centre at the
University of Ottawa Heart Institute
New Microchip Prototype
Testing Currently Underway
Dr. T. Mussivand, FRSC
Medical Devices Centre at the
University of Ottawa Heart Institute
Overall Device Status
– 15 minute Extraction (Original Target):
• Rapid sampling & extraction method developed
• 3rd party validation of results by RCMP
• Patents applied for Canada, US, Japan, Europe
(Method and apparatus for collecting cells for
macromolecular analysis)
– 15 second Extraction (Current Target):
• Preliminary development of rapid cell membrane
lysing technique suitable for a portable device
• Microchip prototype developed and system testing
underway
• Additional work underway on overall device
integration and prototype development
15
Dr. T. Mussivand, FRSC
Medical Devices Centre at the
University of Ottawa Heart Institute
The DNA/Telemedicine Connection
Wednesday March 12, 2008
•
Original Concept: Needed security method for
artificial heart telemedicine system
•
DNA seemed to be the most secure approach
•
The importance of telemedicine security is
highlighted in recent media reports that outline
successful hacking of implantable medical devices
including the ability to:
– Deliver ICD shocks, or
– Turn off the device
•
This work entitled “Pacemakers and Implantable
Cardiac Defibrillators: Software Radio Attacks and
Zero-Power Defenses” will be presented at the
upcoming 2008 IEEE Symposium on Security and
Privacy in Berkeley in May
– The publication was released early online at:
www.secure-medicine.org
Dr. T. Mussivand, FRSC
Medical Devices Centre at the
University of Ottawa Heart Institute
Specific Telemedicine Examples
for a Portable DNA Extraction Device
• Telemedicine Security: Prevent unauthorized access to
telemedicine systems, including for control/monitoring of life critical
devices.
• Other BioSecurity: DNA is considered by some to be the “holy
grail” of biometrics. Data could be transferred for comparisons to
central identity databases. Biosecurity apps include everything from
secure facilities access to transportation safety.
• Population Mapping: The ability to map large communities for
specific genetic variants associated with various health conditions
could be accomplished easily with a portable DNA sampling and
extraction device. Data could be transferred from the field in realtime to international research centres.
Dr. T. Mussivand, FRSC
Medical Devices Centre at the
University of Ottawa Heart Institute
Summary & Conclusions
• New techniques for rapid (15 minute) DNA sampling
and extraction have been developed and validated
• The ability to obtain DNA from both invasive and
non-invasive (fingerprint) samples has also been
demonstrated
• Work is proceeding for a 15 second DNA extraction
system utilizing microchips for a portable device
based system
• Microfluidics based cell membrane lysis (6 seconds)
has been demonstrated at the microchip level
• A wide variety of potential applications in medical
and non-medical fields are possible
• Utilization of the DNA system could provide vital
security for life critical telemedicine applications
• Further work is currently underway
Dr. T. Mussivand, FRSC
Medical Devices Centre at the
University of Ottawa Heart Institute