floating remains: a globalfiler® case study
TRANSCRIPT
Floating Remains: A GlobalfilerTM Case Study
Steen Hartsen Forensic Science & Technology
Financial Disclosure
• I have no financial involvement with Life Technologies or Thermo-Fisher products that will be discussed.
• Life Technologies has sponsored me to be here to speak today.
BCIT Forensics
• Casework
• Education
• Research
Jason Moore - QA
Dean Hildebrand – Technical Leader
• Casework
– British Columbia Coroner Service • Unidentified Human Remains
– Insurance Corporation of British Columbia • Fraud Investigations
– National Forensic Science Institute (Namibia) • Murder/Sexual Assault cases
– Private Clients • Highly variable
BCIT Forensics
• Education
– Three main areas of forensic study: Forensic Science, Computer Crime and Crime & Intelligence Analysis.
– Workshops for various law enforcement agencies on a “as-needed” basis.
– Forensic Biology: DNA Typing Applications
– High School CSI Academy
BCIT Forensics
• Research
– Graduation Research Projects
– Directed Volunteer Projects
– Practicum Placements
– Validation Studies
BCIT Forensics
- Airbags as Sources of DNA Evidence from Motor Vehicle Incident Reconstructions
- A Novel Approach to Cast-Off DNA Analysis Using Globalfiler Express
- Evaluating the Effect of Globalfiler on Familial Searches
Grad Projects:
BCIT – Validation Timeline
2013: GlobalFiler Casework validated
2013: GlobalFiler Casework released
2012: Identifiler
Plus validated
2010: Identifiler
Plus released
2007: Minifiler validated
2007: Minifiler released
2004: Quantifiler
2000: Casework at BCIT
1997: Profiler
Plus released
Case Study: Bridge Jumper
• Deceased was last seen jumping from the Port Mann Bridge over the Fraser river on May 6th, 2012.
• Jumper was a 6’3” tall male.
• The body was not recovered.
Case Study: Bridge Jumper
• March 1st 2013, a left femur is found on the bank of the Fraser River. (PMI: 10 months)
• Anthropologists estimated the height of the individual to be over 6’0”.
• GIS (Geographic Information System) was used to narrow the scope of possible victims
Case Study: Bridge Jumper
• A full profile was obtained with Globalfiler at 1 ng of input DNA.
• Comparisons to family members of the deceased were able to confirm the identity of the jumper.
• The profile was placed in the BCCS database in case any other body parts are found over time.
Case Study: Bridge Jumper
Case Study: Lost Lagoon Floater
Lost Lagoon Floater
• June 6th 1972, a body is discovered by a jogger.
• Individual was determined to be Caucasian. The only identifying features were black pants and shoes.
• No-one was reported missing that matched that description.
Lost Lagoon Floater
• After the autopsy, the remains were buried with the identity undetermined.
Lost Lagoon Floater
• New information came to light in 2013 that allowed investigators to narrow down the identity of the UHR.
• An East Indian individual had been reported missing in April of 1972. The estimated PMI of the remains was about 1 month, indicating it might be a match.
• The missing individual also had black shoes..
Lost Lagoon Floater
• The body had to be exhumed, as no samples had been preserved for future testing.
• During the exhumation, we discovered that the body had been buried in a particularly wet portion of the cemetery.
Sample Processing
• The femur was processed using an organic extraction method and then quantified with Quantifiler Human (35 ng recovered).
• Identifiler Plus was initially used for STR analysis.
• A partial profile was obtained, with data at 14 loci above our analytical threshold of 50 RFU (ABI 310)
Globalfiler Analysis
• Globalfiler validation began the following week on our new 3500 Genetic Analyzer
• We used various types of casework samples to
determine our analytical threshold and sensitivity study, and thought this would be a great test case.
310 Genetic Analyzer
3500 Genetic Analyzer
Reproducibility and Precision of Allelic Ladder
Dye Average StDev N Min RFU Max RFU Avg + 10 SD (LOQ) Analytical ThresholdBlue 14.3 7.2 3880 2 159 86 150Green 7.5 7.1 3834 1 134 78 150Yellow 7.3 4.2 3734 1 80 49 100Red 10.1 8.1 3968 2 221 92 150Purple 10.0 4.6 4096 2 75 56 100
Setting the Analytical Thresholds • 10x 1ng samples, 3x 0.5 ng samples and 4x NAC • Allele peaks and artifacts (pull-up or stutter) were removed from the study as
they are not true noise
Peaks N Total Noise Peaks % noise/total Total Samples % noise/sampleabove LOQ 28 19512 0.14% 17 165%above AT 4 19512 0.02% 17 24%
Sensitivity
Stochastic Threshold GlobalFiler: False Homozygote Call Potential Percentage of False Homozygous Calls
29.8% 7.4% 1.1% 0%
ST=200 RFU ST=300 RFU ST=400 RFU ST=450 RFU
Stutter SE33
N+4 Stutter N-3 & N+3 Stutter N-4 & N-2 Stutter
3.7% 8.9% 2.9% 7.0% 3.7%
9947a Control Heat Map
9947a D3S135
8
vWA
D16S53
9
CSF1PO
TPOX
Yind
el
AMEL
D8S117
9
D21S11
D18S51
DYS391
D2S441
D19S43
3
TH01
FGA
D22S10
45
D5S818
D13S31
7
D7S820
SE33
D10S12
48
D1S165
6
D12S39
1
D2S133
8
rpt 11 ng rpt 2
rpt 3rpt 1
0.5 ng rpt 2rpt 3rpt 1 132
0.25 ng rpt 2 147rpt 3rpt 1
0.125 ng rpt 2rpt 3rpt 1
0.064 ng rpt 2 246rpt 3rpt 1 155 272
0.032 ng rpt 2 321 162 235 321 311rpt 3 315 207 247 364 297 316 256rpt 1 215 173 147 113
0.016 ng rpt 2 113 151rpt 3 217 153 112
Back to our Floater..
Bone D3S135
8
vWA
D16S53
9
CSF1PO
TPOX
Yind
el
AMEL
D8S117
9
D21S11
D18S51
DYS391
D2S441
D19S43
3
TH01
FGA
D22S10
45
D5S818
D13S31
7
D7S820
SE33
D10S12
48
D1S165
6
D12S39
1
D2S133
8
rpt 11 ng rpt 2 198
rpt 3 165rpt 1 335
0.5 ng rpt 2rpt 3 256 144 172rpt 1 200 179 215 140 101
0.25 ng rpt 2rpt 3rpt 1
0.125 ng rpt 2 242 151 119rpt 3 257 260 162 119rpt 1 162 156
0.064 ng rpt 2 117rpt 3 131 188rpt 1 187
0.032 ng rpt 2 158rpt 3rpt 1 105
0.016 ng rpt 2rpt 3
Back to our Floater..
Back to our Floater..
• The profile obtained was compared to a surviving family member.
• Familial comparison proved it was indeed a match
• Remains were cremated at the family’s request.
Project Amplify
• Student project focusing on retrospective amplification for unresolved cases within the BC Coroner Service.
• Examining 26 outstanding cases to try and expand the database entries
As of August 2012, 44 countries had active national offender DNA databases. www.dnaresource.com
DNA Results from BCCS Samples (2012)
Project Amplify
• Most cases have low statistical resolvability when making familial comparisons
• Typically they have been amplified with Profiler Plus.
• As the DNA database has grown, fortuitous matches have increased due to limited loci.
Probability of Identity
Profiler Plus CODIS Identifiler Plus GlobalFilerLoci 9 13 15 21Prob of Identity 1.48E-‐11 2.13E-‐15 5.01E-‐18 3.71E-‐26LR 6.76E+10 4.69E+14 2.00E+17 2.70E+251in 67 Billion 469 Trillion 200 Quadrillion 27 Septillion
Project Amplify
• Increasing the number of loci increases the stats
• Direct comparisons go from astronomical to even more astronomical
• Familial comparisons can go from moderate to very strong support.
LR: 1 in 2798
Source: “EASY DNA” by Dr. Fung
LR: 1 in 841 Thousand
Source: “EASY DNA” by Dr. Fung
LR: 1 in 241 Million
Source: “EASY DNA” by Dr. Fung
Project Amplify
• Work continues to update unresolved cases with as much new data as possible.
• I would like to thank the BC Coroner Service: Identification & Disaster Response Unit for allowing me to talk about these cases.
