Direct Amp Comparison of Multiplex Kits on Challenging DB SamplesBRITTANY ROSS

KENTUCKY STATE POLICE CENTRAL FORENSIC LABORATORY

New STR Kit Challenges

Fall 2018 CODIS Conference

Joint presentation by Brian Harmon and Lisa Grossweiler, “Artifacts and Oddities – New STR Kits” Issues the CODIS community are seeing with GlobalFiler, Fusion &

Investigator 24plex kits

Issues that impact both Database & Casework Non-concordances between kits, even older generations of the

same company’s kits

Marker Range Issues: Does this allele belong to D1 or D12?

Migration & 1bp Resolution Issues

Recommended solutions Validation of a SECONDARY STR kit

Validation of a Secondary Kit

Validation of a Secondary Kit

Express

KSP DNA Database History

KSP Database running Identifiler Plus Average annual samples received:

15,000 2013 received almost 25,000 due to a

DOC catch-up

Sample format: buccal swabs transferred to FTA indicating cards

Post-2008 2013-2015 2016 2017

KSP DNA Database History

2013: Expanded loci kit debate begins! Promega Fusion 5C vs AB GlobalFiler/Express Summer 2013: AB offers to “validate” new

DB 3500 as a way to let us evaluate GlobalFiler Express

2014: continued evaluations for 5C & GF 2015: selection & validation begins

Post-2008 2013-2015 2016 2017

KSP DNA Database History

March: all processing ceased to allow for 5C training & competencies

April 1st: processing in 5C began Year in review:

Backlog had swelled to 19,440 offender samples by April

13,911 samples received in 2016

Samples processed: 29,727

Post-2008 2013-2015 2016 2017

KSP DNA Database History

Decision made to bring another kit online

Previous 2013 AB GFE validation work was incomplete Did not include work with older sample formats (blood on

S&S Whatman 903 paper)

A 2nd 3500 & 2 Veriti thermocyclers purchased in 2016

Supplemental validation completed that spring

GFE online by August Qiagen approached us about 24plex GO!, have

started validation

Post-2008 2013-2015 2016 2017

Challenging Samples

Vast majority of problem samples are weak Sometimes just d/o at TPOX and/or D22 in 5C

Marker overlap Most often between D2S441/D10 & D12/D19 for 5C

DYS391 issues

Null Markers or alleles D12 migration issues

Blobs

Sample 1: Blobs - Fusion

Amp 1 Amp 2

Hiding 3rd allele?(unlikely)

Sample 1: BlobsGFE GO!

Truly homozygote?

Sample 2: Blobs - Fusion

Amp 1 Amp 2

Interferes w/13 Can’t discern 13

Sample 2: Blobs

GFE GO!

Sample 3: Null D19

GO!

GFEID

5C

Sample 4: Null D22

5C

GFEGO!

Sample 4: Null DYS570

6C

Y23

Yfiler Plus

Sample 5: Null vWA allele

5C

GFE

GO!

ID

Sample 6: DYS391 non-concordance

6C

5C

GO!

10.3469.2bp

GFE

AL 11470.3bp

Sample 6: DYS391 non-concordance

Y23Typed the same

as 6C/GFE

Yfiler PlusTyped the same

as 5C

10.3376.3bp

AL 11377.3bp

Sample 7: DYS391 non-concordance

5C

GFE

GO!

8.3372.5bp

AL 9373.5bp

Sample 7: DYS391 non-concordance

Y23Typed the same as 5C

Yfiler PlusTyped the same as GFE

D12 migration

Sample 8: D12 migration

GFE GO!5C 3130

OL 165.4bp

21.2 246.5bp

AL 21 244.5bp22 248.6bp

2nd amp, 2nd Inj

OL 164.9bp

AL 21 162.9bp22 167.2bp

AL 21 162.5bp22 166.6bp

2nd amp, 1st Inj

5C 3500

21.2 233.8bp

AL 21 231.8bp22 235.9bp

Sample 9: D12 migration

GFE GO!

OL 167.4bp

22 248.5bp

AL 22 248.6bp

2nd Inj

OL 167.4bp

AL 22 166.7bp

AL 22 166.9bp

1st Inj

5C

22 236.1bp

AL 22 236.1bp

Sample 10: Marker Cross-Over

6C

GFE

GO!

5C

Sample 11: Marker Cross-Over

GFEGO!

5C

Sample 12: Marker Cross-Over

5C

GFE

GO!

ID

Sample 13: Marker Cross-Over

5C

GFE

GO!

ID

Acknowledgements

KSP Database Shannon Keeton Danielle Kuntz Nick Zimmerman Regina Wells

Contact Info

Brittany Rossbrittany.ross@ky.gov