GlobalFiler Kit Casework Test Site Results and Other Development Updates April Orbison Sr. Field Applications Specialist Human Identification 1
Agenda GlobalFiler Kit Update: Recent Casework Test Site Results Development Updates Next Generation Y-STR (NGY) Multiplex Next Generation Quantification Systems HID Workflow Software Solutions ParaDNA Screening System Sample Collection 2
What is the GlobalFiler Kit? Locus CODIS (Current) Europe CODIS (Proposed) GlobalFiler Kit D13S317 X X X D7S820 X X X D5S818 X X X CSF1PO X X X D1S1656 X X X D12S391 X X X D2S441 X X X D10S1248 X X X D18S51 X X X X FGA X X X X D21S11 X X X X D8S1179 X X X X The GlobalFiler Kits are our response to the worldwide expansion of forensic locus sets, recently captured in the CODIS Core Loci Working Group s recommendation to expand the CODIS Core Loci from 13 to 20 required loci and 3 highly recommended loci VWA X X X X D16S539 X X X X TH01 X X X X D3S1358 X X X X AMEL X X X X D2S1338 X X X X D19S433 X X X X DYS391 X X TPOX X X X D22S1045 X X X 1. D.R. Hares, Expanding the CODIS core loci in the United States, Forensic Sci. Int. Genet. 6 (2012), e52-e54. 2. D.R. Hares, Addendum to expanding the CODIS core loci in the United States, Forensic Sci. Int. Genet. 6 (2012), e135. SE33 3 X X X
The GlobalFiler Kits: 6-Dye Configuration 1 Y indel Amel 3 2 11 12 13 14 DYS391 Only 2 loci require primer redesign 18 Mini-STRs Combination PI*= 5.73 x 10-21 4 7 5 6 8 16 17 15 All gender markers located in the same dye channel for ease of interpretation 9 10 18 GlobalFiler Kit 10 Mini-STRs PI* = 9.2 x 10-12 MiniFiler Kit 8 Mini-STRs PI* = 1.05 x 10-10 97% of alleles in allelic ladder <400bp 4 * Preliminary calculations based on a US Caucasian Database
The GlobalFiler Kits: Allelic Ladder 31 alleles added to 9 GlobalFiler Kit marker ladders New virtual bins added to improve genotyping efficiency 343 alleles 246 virtual bins 589 total alleles 5
SE33 Locus High Number of Alleles & Virtual Bins 6
Facts about the SE33 Locus The most discrimination power of any STR locus in commercial kits highest Het obs (0.9353) lowest P I value (0.0066) most amount of alleles and genotypes observed Ideal for mixtures more alleles seen and less allele sharing Widely Used in Europe Included in the Applied Biosystems SEFiler, SEfiler Plus and NGM SElect kits for many years Becoming more widely used internationally - Hill, B et al, "Population Statistics on the Proposed Expanded U.S. Core Loci", poster at the 23rd International Symposium on Human Identification (ISHI) meeting (Nashville, TN), October 16-17, 2012 7
SE33 Locus Ideal for Mixtures 8
Maximizing Discrimination Kit D8S1179 D21S11 D7S820 CSF1PO D3S1358 TH01 D13S317 D16S539 D2S1338 D19S433 vwa TPOX D18S51 D5S818 FGA D10S1248 D22S1045 D2S441 D1S1656 D12S391 SE33 PI Identifiler Kit X X X X X X X X X X X X X X X 5.01 x 10-18 NGM SElect Kit X X X X X X X X X X X X X X X X 2.35 x 10-21 GlobalFiler Kit X X X X X X X X X X X X X X X X X X X X X *7.12 x 10-26 GlobalFiler Mini s X X X X X X X X X X *9.20 x 10-12 * Preliminary calculations based on a US Caucasian Database The GlobalFiler Kits will: Deliver a new level of discrimination Reduce the risk of adventitious matches Facilitate international DNA profile comparisons 9
GlobalFiler Kit: Casework Design Considerations Casework Kit Performance Requirements: High sensitivity for trace/low level DNA samples Good intracolor balance for mixture samples Sufficient tolerance to PCR inhibitors Maintain high species specificity Efficient thermal cycling conditions 10
Optimization of GlobalFiler Kit Reaction Conditions Reagent Volumes 3.3X Master Mix 7.5 µl 10X Primer Set 2.5 µl Sample Input 15.0 µl 25.0 µl PCR Conditions 95 C/ 1 min 94 C/ 20 sec 59 C/ 90 sec 60 C/ 10 min 29 x Greater flexibility of sample input volume to facilitate analysis of low level samples Optimized thermal cycling conditions shorten amplification conditions by > 1 hr Identifiler Plus & NGM SElect Kits GlobalFiler Kit Total Time 2 hr 25 min 1 hr 20 min 11
GlobalFiler Kit Performance Example: Sensitivity [DNA] 1 ng 500 pg 250 pg 125 pg 62.5 pg 31.25 pg 15.63 pg GlobalFiler Kit Yind SE33 DY3 D22 D2(S4) D12 D10 D1 FGA D5 Amel D18 TPOX vwa D19 D2(S1) D16 D13 TH01 D3 CSF D7 D21 D8 Identifiler Plus FGA D5 Amel D18 TPOX vwa D19 D2 D16 D13 TH01 D3 CSF D7 D21 D8 Greater sensitivity was observed with the GlobalFiler Kit compared to the Identifiler Plus Kit The GlobalFiler Kit exhibits slightly greater sensitivity than the Identifiler Plus Kit 12
GlobalFiler Kit Performance Example: Inhibitor Tolerance Humic Acid Inhibition Hematin Inhibition Identifiler Plus Kit GlobalFiler Kit The GlobalFiler Kit buffer has been optimized to deliver enhanced inhibition tolerance compared to the Identifiler Plus Kit 13
GlobalFiler Kit Test Site Data 16
GlobalFiler Kit Test Site Overview TS1 TS2 TS3 TS4 TS5 Extraction Method Organic or EZ1 Organic Magnetic Bead Qiagen Investigator Organic Comparative Kit Identifiler Plus, PP 16HS, PP Fusion NGM SElect Identifiler Plus Identifiler Plus NGM SElect Thermal Cycler 9700 Gold Veriti Veriti 9700 Gold 9700 Gold CE Instrument 3500 3500 3500xL 3500xL 3500 17
GlobalFiler Kit Baseline Quality 74 444 Read Region 300 Test Site #1 300 Test Site #2 300 Test Site #3 Non-template Controls 18
GlobalFiler Kit Performance Example: Sensitivity GlobalFiler Kit [DNA] D8 D21 D7 CSF D3 TH01 D13 D16 D2(S1) D19 vwa TPOX D18 Amel D5 FGA D1 D10 D12 D2(S4) D22 DY3 SE33 Yind 1 ng 500 pg 250 pg 125 pg 62.5 pg 31.25 pg 15.63 pg Increased allele recovery with low input samples. 19
Alleles Recovered Using the GlobalFiler and Identifiler Plus Kits : Partial Profiles 45 40 35 No. Alleles Recovered 30 25 20 15 GF Partial ID+ Partial 10 5 0 Fat Trace Swab /310pg Key Swab/ 60pg Bottleneck Swab / 50pg Drawer Swab/60pg Swab on Safe / 50pg Wheel Swab /80pg Partial Profile Samples Cigarette Butt / 40pg Cigarette Butt / 30pg Cap Swab / 165 pg Stone / Undet The increased robustness of the GlobalFiler Kit delivered 182% more alleles than Identifiler Plus with partial profiles Deliver more information per sample Increases the likelihood of a meaningful database match 23
GlobalFiler Kit Test Site Results Identifiler Plus Kit: Full (29 alleles) GlobalFiler Kit: Full (40 alleles) Bone Sample; Magnetic Bead Extraction (1ng) Identifiler Plus Kit: 50 RFU PAT (3130xl) GlobalFiler Kit: 175 RFU PAT (3500xL) 26
GlobalFiler Kit Test Site Results Identifiler Plus Kit: Partial Profile (21 alleles) GlobalFiler Kit: Partial Profile (28 alleles) Bone Sample; Organic Extraction Identifiler Plus Kit: 50 RFU PAT (3130xl) GlobalFiler Kit: 175 RFU PAT (3500xL) 27
GlobalFiler Kit Test Site Results Identifiler Plus Kit: Partial (21 alleles) GlobalFiler Kit: Full (40 alleles) Door Handle Swab, Qiagen Investigator (310pg) Identifiler Plus Kit: 50 RFU PAT (3130xl) GlobalFiler Kit:175 RFU PAT (3500xL) 28
GlobalFiler Kit Test Site Results PowerPlex Fusion Kit: Partial (33 alleles) GlobalFiler Kit: Full (42 alleles) Sample Type: Unknown GlobalFiler Kit: 175 RFU PAT (3500) PowerPlex Fusion:175 RFU PAT (3500) 31
Development of a Next Generation Global Multiplex Go Faster! Up to 5x faster amplification time than previous generation kits Go Further! > GlobalFiler Express Kit: ~40 minutes; GlobalFiler Kit: ~80 minutes > Enables sample to answer in <2 hours 24 loci, including 10 mini-strs, increasing discrimination power by up to 9 orders of magnitude Enhanced buffer system and sensitivity for even better performance on inhibited and low-level samples Go Global! The only kit containing all markers recommended for inclusion by the CODIS Core Loci Working Group, including all markers commonly used in Europe Maximizes global database compatibility 34
Yfiler Plus Kit 35
Development of a Next Generation Y-STR Multiplex Designed to enable: Increased discrimination power and potential to discriminate between close male relatives with expanded marker set Higher number of alleles recovered at low DNA input amounts Improved recovery of alleles and intracolor balance in male/female mixtures containing high female background Improved resolution capacity for male/male mixtures Improved recovery of alleles in inhibited samples 36
Yfiler Plus Kit Configuration 50 100 150 200 250 300 350 400 DYS627 DYS389II DYS635 DYS389I DYS576 1 2 DYS391 DYS448 Y GATA H4 DYS19 DYS458 DYS460 3 4 5 DYS518 DYS392 DYS438 DYS390 DYS456 6 7 DYS449 DYS385ab DYS437 DYS570 8 9 DYS533 DYF387S1ab DYS481 DYS439 DYS393 New Y-STR Markers 37 10 11 Rapidly Mutating Y- STR Standard Y-STR
Web based Y-STR resource hub and information center developed to: Y-STR Community Provide the most updated information in the area of forensic Y-STR analysis Facilitate communication between Life and its customers Help forensic scientists around the world collaborate and stay in touch http://www.lifetechnologies.com/ycommunity 38
Next Generation Quantification 39
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Next Gen Quantifiler Human & Quantifiler Duo Kits More Accurate, Sensitive, and Specific Take Control of your Workflow Fall 2013 Fast & Accurate Decision Tool Shorter time to results as compared to original quantification kits Increased assay sensitivity and accuracy Solid decision making for forensic casework sample processing pathways Higher Performance with Challenging Samples Improved performance with low-ratio male/female mixture samples IPC optimization to match performance of next generation STR kits Gain key information on sample quality prior to PCR amplification Enhanced Workflow & Data Analysis Improved consistency and homogeneity of DNA standard Customizable quantification workflows for complex casework sample processing Two new Quantifiler Kits in 2013 To integrate with Next Generation STR kits 41
Prototype Sensitivity Data Consistency 1.2 pg/µl 20 ng/µl DNA (ng/µl) 42 The content provided herein may relate to products that have not been officially released and is subject to change without notice
Prototype Sensitivity Data Consistency 1.2 pg/µl 20 ng/µl DNA (ng/µl) 43 The content provided herein may relate to products that have not been officially released and is subject to change without notice
Be Part of the Change! Become a Test Site Help Us Grow the Population Databases Support our rigorous development process Give us feedback and help us build the kits that meet your needs Be the first to evaluate the NGY and Quantification kits with your challenging samples Become part of the Next Generation Y-STR Multiplex Community Help other forensic laboratories by sharing your population data Contact your local Human Identification support and sales representative 2013 Life Technologies Corporation. All rights reserved. The trademarks mentioned herein are the property of Life Technologies Corporation and/or its affiliate(s) or their respective owners. For Forensic or Paternity Use Only. 44
Converge Software Solutions 45
Current Software Offering in Life Tech HID workflow Case and Sample Information Mgmt DNA Workflow Management Genotype Profile Generation Genotype Profile Storage Genotype Profile Analysis Tools Software data management tools in Real-time QPCR Quantification (HID Real-Time and SDS 1.2.3) Fragment analysis data collection CE software (UDC and MDC) Expert system genotyping software (Genemapper ID-X) 46
Feedback we have heard from customers We waste a lot of time searching for data through multiple systems for reviews and audits The DNA workflow steps are manually controlled and tracked We need easier and flexible systems for kinship analysis Its tedious and costly to store data and storage can be unreliable We want to be able to easily build and search local forensic databases I hate having to enter sample information multiple times in the different systems used in the DNA workflow We want to be able to have tools to support familial search application 47
How can we better integrate your DNA workflow from case to report Case and Sample Information Mgmt DNA Workflow Information Management Genotype Profile Generation Genotype Profile Storage Genotype Profile Analysis Tools Centralized data storage, access and instrument integration Converge DNA Workflow Integrate your DNA workflow instrumentation, worksheets, data storage and access Converge Kinship and Paternity Simplify your case management, analysis and reporting Converge Direct Search Build and search your own local profile databases using direct matching methods Converge Familial Search Build and search your own local profile databases using a novel familial search algorithm 48
Modularity and Customizability is Key to Converge Modular software development platform for HID labs providing tools for: Kinship Analysis Local Database Generation Local database searching (direct and familial) Data Management Systems integration Workflow management Modules can be purchased individually or together in bundles. Modules integrate together and with existing HID software such as GeneMapper ID-X Life Technologies Quantification Software (HID Real-Time and SDS 1.2.3) Life Technologies Data Collection Software (MDC and UDC) Heavy configuration to meet your labs specific workflow and requirements First set of modules presented here will be released in mid year 2013 49
ParaDNA Screening System 54
Understand More about the Quality of Human DNA in Your Evidentiary Samples Available Now Sample to Answer Casework Screening Device used by law enforcement agencies prior to evidence submission to the crime laboratory Screening Unit Screening Tests ACTIONABLE RESULTS for law enforcement decision making to support EVIDENCE PRIORITIZATION and case investigation process MAXIMIZE EFFICIENCY of sample submission and MINIMIZE chances for evidence cross CONTAMINATION SAMPLE TO ANSWER FORMAT 55 Sample Collector
Nicola Oldroyd Consultant Forensic Scientist Life Technologies Sample Collection Updates Reference and Crime Scene DNA Collection 56
NUCLEIC-CARD System-Now Available in the US Scheduled to A reliable device for collection, preservation, and long-term storage of nucleic acids NUCLEIC CARD NUCLEIC CARD Collection Device NUCLEIC CARD COLOR Chemically-treated matrix to enable cell lysis and protein denaturation 57
NUCLEIC-CARD Collection Device Easy-to-use sample collection device for buccal cell collection Better Collection Lollipop-shaped 4N6FLOQSwabs Higher Transfer Efficiency Uniform and reproducible sample transfer Easier Processing Framed NUCLEIC-CARD suitable for automatic punching systems 58
Introducing the 4N6FLOQSwabs Excellent recovery of DNA Maximum efficiency in collection capacity Available with different anatomical and ergonomic design Human DNA, Dnase and RNase-free certified ETO-treated (Ethylene Oxide) Pre-scored breaking point Flocked Swab Patented technology to maximize DNA collection and elution efficiency 59
4N6FLOQSwabs for Crime Scene Anatomically and ergonomically shaped Optimized for challenging samples Treated with antimicrobial agents No need for drying 60
Web & Event Resources 61
www.lifetech.com/ forensicnews 62
HID University Seminar Series 2013 Dates & Locations: Washington D.C. - Tuesday. July 9 th Crystal City Double Tree Phoenix, AZ Tuesday. July 16 th Montelucia Chicago, IL Thursday, Aug 6 th Marriott Chicago Downtown Oakland, CA Tuesday, Aug 13th Marriott City Center Atlanta, GA Thursday, Aug 15 th InterContinental Buckhead Key Themes & Topics: The GlobalFiler Kit: Going Further for Casework Next Generation Y-STR and Quantification Assay Development Updates Workflow software solutions and efficiency improvements Tools for improving DNA collection & preservation Casework screening solutions Global legislative updates Ion Torrent Semiconductor Sequencing for Forensic Applications Interesting Cases Check for updates on the HID University Registration Page: www.lifetechnologies.com/hiduniversity 63
For more information: www.lifetech.com/hid www.slideshare.net/lifetech_hid Thank You!! 2013 Life Technologies Corporation. All rights reserved. The trademarks mentioned herein are the property of Life Technologies Corporation or their respective owners. The content provided herein may relate to products that have not been officially released and is subject to change without notice. FOR FORENSIC AND PATERNITY USE ONLY 64