1. © Forensic Science Service Ltd. 2010. All rights reserved. New DNA developments better, faster and cheaper Martin Bill, R&D Forensic Science Service Ltd SPSA / SIPR conference 14 th & 15 th September

2. © Forensic Science Service Ltd. 2010. All rights reserved. Agenda 1.Better – new multiplexes 2.Faster – Custody suite DNA testing 3.Cheaper – DNA INSIGHT

3. © Forensic Science Service Ltd. 2010. All rights reserved. Better – new multiplexes Assessing the performance of the new miniSTR chemistries

4. © Forensic Science Service Ltd. 2010. All rights reserved. ENFSI Requirement Robust multiplexes, inc. Mini STRs Fifteen STR loci –Ten SGM Plus ® loci, FGA, TH01, VWA, D3S1358, D8S1179, D18S51, D21S11, D2S1338, D16S539, D19S433 –Additional 5 loci D1S1656, D2S441, D10S1248, D12S391, D22S1045 –Amelogenin ENFSI DNAWG December 2009 –Expansion of ESS to include additional 5 loci.

5. © Forensic Science Service Ltd. 2010. All rights reserved. Strategic multiplexes Two different approaches –Make SGM Plus ® Markers Standard –Make SGM Plus ® Markers Mini Response Size /bp

6. © Forensic Science Service Ltd. 2010. All rights reserved. Strategic multiplexing Choice of PowerPlex® multiplexes ESX-16 & ESX-17 – Standard size SGMPlus ® loci ESI-16 & ESI-17 – Mini SGMPlus ® loci Optimised for 500pg DNA and 30 cycles Applied Biosystems NGM BioType Mentype® ESSplex

7. © Forensic Science Service Ltd. 2010. All rights reserved. ESX D2S1338D16S539D1S1656D10S1248D19S433D2S441 D12S391 D22S1045 vWA D8S1179FGA SE33 Amel D3S1358 D18S51 D21S11 TH01

8. © Forensic Science Service Ltd. 2010. All rights reserved. ESI Amel D3S1358 D19S433D2S1338D22S1045 D16S539D18S51D1S1656D10S1248D2S441 D8S1179FGA SE33 vWAD21S11D12S391TH01

9. © Forensic Science Service Ltd. 2010. All rights reserved. Evaluation of Performance Sensitivity Inhibitors Degraded samples Concordance

10. © Forensic Science Service Ltd. 2010. All rights reserved. Sensitivity

11. © Forensic Science Service Ltd. 2010. All rights reserved. Sensitivity

12. © Forensic Science Service Ltd. 2010. All rights reserved. Sensitivity DNA Template (pg) 1 locus 10 loci

13. © Forensic Science Service Ltd. 2010. All rights reserved. Working Range

14. © Forensic Science Service Ltd. 2010. All rights reserved. Inhibitors Humic Acid Indigo Haematin

15. © Forensic Science Service Ltd. 2010. All rights reserved. Humic Acid

16. © Forensic Science Service Ltd. 2010. All rights reserved. Indigo

17. © Forensic Science Service Ltd. 2010. All rights reserved. Haematin

18. © Forensic Science Service Ltd. 2010. All rights reserved. Degraded DNA ESX vs ESI vs SGMPlus®

19. © Forensic Science Service Ltd. 2010. All rights reserved. Analysis of degraded DNA SGMPlus ® with degraded DNA

20. © Forensic Science Service Ltd. 2010. All rights reserved. Degraded DNA ESX-16 with degraded DNA

21. © Forensic Science Service Ltd. 2010. All rights reserved. Degraded DNA ESI-16 with degraded DNA

22. © Forensic Science Service Ltd. 2010. All rights reserved. Degraded DNA SampleESXESISGM+ 16 loci11 loci16 loci11 loci Degraded 56%38%63%86%33% FSS UV138%27%34%50%0% FSS UV216%9%28%41%ND ENZ 1065%64%65%86%59% ENZ 1542%36%58%77%36% UV semND 100% 76%

23. © Forensic Science Service Ltd. 2010. All rights reserved. Degradation

24. © Forensic Science Service Ltd. 2010. All rights reserved. Concordance NIST FSS

25. © Forensic Science Service Ltd. 2010. All rights reserved. NIST Concordance with Identifiler ® 1461 samples ESX-16 –4 loci discordant –99.98% concordance ESI-16 –3 loci discordant –99.98% concordant

26. © Forensic Science Service Ltd. 2010. All rights reserved. FSS Concordance with SGMPlus ® Number of loci compared Number of concordant loci Concordance (%) ESX 163500349899.94 ESI 163500349999.97 Non-concordant alleles currently being sequenced ~1 in 200 profiles non-concordant

27. © Forensic Science Service Ltd. 2010. All rights reserved. Summary Promega ES multiplexes –Sensitive –Robust to inhibitors –ESI SGMPlus loci very robust with degraded DNA –High level of concordance with NIST samples

28. © Forensic Science Service Ltd. 2010. All rights reserved. Next steps: For new multiplexes meeting the ENFSI requirement: –Gain acceptance to NDNAD –Concordance (ESX vs ESI vs SGMPlus® vs NGM vs ESSplex) –Frequency databases –Check for independence (product rule)

29. © Forensic Science Service Ltd. 2010. All rights reserved. Development of a forensically integrated DNA analysis system. Faster - Custody suite DNA testing

30. © Forensic Science Service Ltd. 2010. All rights reserved. Introduction What is the aim? What have we accomplished What will we achieve in the future

31. © Forensic Science Service Ltd. 2010. All rights reserved. Suspect Arrest DNA sample taken Submissio n process DNA lab Profile interp DNA loaded to NDNAD Current Process Suspect Released 6-7 h Match

32. © Forensic Science Service Ltd. 2010. All rights reserved. Requirement Environment –deployment Operator –Skill –Training Outcome –DNA profile loaded to the NDNAD –Match reported in Real Time Robust Simple User Interface Automated –Chemistry –Analysis –Database search 2h turn round Evidential quality Eliminate submission by processing at POC

33. © Forensic Science Service Ltd. 2010. All rights reserved. What weve accomplished 4h process

34. © Forensic Science Service Ltd. 2010. All rights reserved. How does it work? Chambers Channels Pumps Valves Vents ECB

35. © Forensic Science Service Ltd. 2010. All rights reserved. PCR Chamber Direct to µCE Elute Waste V V V V V V VV V V V Wash Magnet Archive Denature

36. © Forensic Science Service Ltd. 2010. All rights reserved. Micro Capillary Electrophoresis 140-mm glass chip PVP/HEC polymer 200 V/cm Detection by LIF λ ex 491 nm

37. © Forensic Science Service Ltd. 2010. All rights reserved. Profile from fully integrated run

38. © Forensic Science Service Ltd. 2010. All rights reserved. Ladders TH01

39. © Forensic Science Service Ltd. 2010. All rights reserved. What we will achieve in the future Single cartridge –Plastic CE Reduce time to <2h Sample to match Multiple samples –Maintain sample integrity Casework samples Integrated lysis vial

40. © Forensic Science Service Ltd. 2010. All rights reserved. Review Simple set up Self contained cartridge Minimised opportunity for contamination; batch control Full 16-plex DNA profiles with no manual intervention Identified requirement for further work (in progress) Software/database solutions essential for benefit realisation

41. © Forensic Science Service Ltd. 2010. All rights reserved. Improved DNA interpretation Cheaper – FSS DNA INSIGHT & DNAboost

42. © Forensic Science Service Ltd. 2010. All rights reserved. 24 26 16 20 How does DNA interpretation work and why is it suboptimal? DNA is visualised as a signal referred to as an EPG. The role of the scientist is to translate the signal into a simple set of paired numbers that define the DNA profile. These numbers are then compared between profiles in a case (suspect to crimestain) or through the National DNA Database. Issue 1: Complex data are impossible to convert into a simple set of paired numbers. ? Issue 2: A high proportion of reporting officer interpretation work is performed manually.

43. © Forensic Science Service Ltd. 2010. All rights reserved. Where does DNAboost fit into all this? DNAboost, in part, addresses Issue 1: Complex data are impossible to convert into a simple set of paired numbers by a simple improvement to the database design. 1517 1518 17 18 15 17 18 Issue 2: A high proportion of reporting officer interpretation work is performed manually remains largely unimproved and has opportunities for improvements in efficiency, response time, effectiveness and quality. DNAboost was always intended to be a development milestone towards improved coding, searching and matching. DNAboost improves the search and match rate for DNA intelligence and demonstrates that improved encoding of data is a key way of improving performance.

44. © Forensic Science Service Ltd. 2010. All rights reserved. How could DNA interpretation be further improved? The current approach always discards useful information that may be required for case assessment, intelligence or troubleshooting. DB National DNA Database or interp process 18, 16, 11 Lost information. Next Generation Interpretation The answer is to develop a new interpretation approach that always retains and uses all the useful information. This requires whole EPG interpretation. Improved capacity & cost effectiveness Needs manual, paper review later in the process. Use all the information in a new continuous model 3 pieces of informatio n. ~80,000 pieces of information. Manual best guess

45. © Forensic Science Service Ltd. 2010. All rights reserved. Design and deployment of the new interpretation model AnalystReporting Officer Interprets batches of DNA data to confirm validity. –Software-based. –Partially automated. –Rule-based & objective Unprocessed digital data Valid digital data Valid paper data Expert opinion Interprets cases of DNA data to form opinion. –Paper-based. –(Very) manual. –Guideline-based and artisan(?)

46. © Forensic Science Service Ltd. 2010. All rights reserved. AnalystReporting Officer –Software to interpret DNA data Settings server FSS DNA INSIGHT APB Data server –Server to regularise and disseminate DNA data –Server to control settings and administration –Science, publication, validation, process, training and support FSS DNA INSIGHT design

47. © Forensic Science Service Ltd. 2010. All rights reserved. Increased capacity

48. © Forensic Science Service Ltd. 2010. All rights reserved. Good examples of forensic science getting better, faster and cheaper… … in reality they all add value in a number of areas

49. © Forensic Science Service Ltd. 2010. All rights reserved. Better, faster, cheaper The new chemistries may increase timeliness through higher first pass success rates with a reduced requirement for specialist methods like LCN Custody suite testing may reduce total forensic supply chain costs. DNA INSIGHT may increase success rates through the improved interpretation models.

50. © Forensic Science Service Ltd. 2010. All rights reserved. Thanks for listening Martin Bill, R&D Forensic Science Service Ltd SPSA / SIPR conference 14 th & 15 th September