floating remains: a globalfiler® case study

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.

Questions?

Email: Steen_Hartsen@bcit.ca