New Technologies for Long-term Preservation and Storage of Forensic DNA Samples

Transcription

1 New Technologies for Long-term Preservation and Storage of Forensic DNA Samples Omo Clement, Ph. D. Director of Marketing AFDAA Summer Meeting, San Antonio, TX. July 8, 2011.

2 DNA instability

3 DNA stabilization/storage is important To maintain sample integrity To re-test samples Critical especially for low quality and low quantity DNA samples (e.g., bone, teeth, touch, etc) Pivotal when sub-optimal conditions may compromise DNA quality high T, chemical contaminants, repeated freeze-thaw, environmental damage, etc.. An efficient long-term DNA storage solution is a key need in forensic DNA analysis.

4 Biomatrica s SampleMatrix formulation SampleMatrix technology is based on principle of anhydrobiosis (life without water) a phenomenon by which nature selfpreserves biological functions Synthetic chemical matrix mimics anhydrobiosis as a dissolvable matrix Formulated for long-term stabilization of DNA (storage and shipping) by forming a protective seal around DNA during the drying process. SampleMatrix Anhydrobiotic organism SampleMatrix Nucleic Acids DNA is preserved in dry matrix until ready to use; simply rehydrate and sample is ready for analysis without further purification.

5 DNA stabilization products Purified gdna extracts DNAstable /QIAsafe DNA (Dry-down & LD) Long-term storage solution DNA in whole blood & buffy coat DNAstable Blood/QIAsafe DNA Blood (Dry-down) Long-term storage solution DNA in Tissue, cells & biopsy materials DNAgard Tissue (Liquid stabilizer) Ideal for field collection & shipping to lab for analysis

6 DNAstable/QIAsafe DNA Dry-matrix format Pre-coated DNAstable tube (or plate) Purified DNA mixed with DNAstable Room Temperature Room Temperature Recover Just add water Liquid-to-dry format (LD) Purified DNA Purified DNA and DNAstable LD Room Temp Room Temp Recover Just add water

7 Long-term DNA storage studies DNAStable DNAstable LD Steve Guroff San Diego Sheriff s Department Study Period: (DNAstable) -20ºC 100ng Ambient Temperature Storage NTC DS 20ng DS 20ng 20ng of purified DNA 3.5 years storage time 500bp Image kindly provided by Steve Guroff, San Diego Sheriff s Department 20 ng of human genomic DNA, stored in DNAstable (DS) for 3.5 years at ambient temperature (RT) were analyzed by PCR amplification using beta-actin specific primers generating a 500 bp amplicon DS = DNAstable NTC = Non-template control 3.5 years at Room Temperature

8 Total DNA (ng) Long-term DNA storage studies Greg Hadinoto LA County Sheriff s Department Study Period: 2006 ongoing (DNAstable) Conventional Freezer Storage (-20 C) versus Room Temperature SampleMatrix Storage 45 Months DNAstable -20C Semen Buccal Bloodstain DNA samples from Blood Buccal swabs Semen Storage time-points 1-month 3-months 6-months 12-months 18-months 24-months 60-months Gregory M. Hadinoto. Evaluation of the Biomatrica SampleMatrix room temperature DNA storage system: 12-month and 45-month data. Poster #60, 21st International Symposium on Human Identification, San Antonio, TX, Oct 11-14, 2010.

9 DNA recovery better than frozen? DNAStable DNAstable LD Crouse, et. al. Palm Beach Sheriff s Office DNAstable /QIAsafe DNA compared to -20C storage 120% 100% 80% 60% 40% 20% 0% 4ng 2ng 1ng 0.5ng 0.25ng 0.125ng ng -20C DNAstable Consistently higher recovery in DNAstable-stored samples Lee, S.B., Clabaugh, K.C., Silva, B., Odigie, K.O., Fourney, R.M., Stevens, J., Carmody, G.R., Coble, M.D., Loreille, O., Scheible, M., Parsons, T.J., Pozder, A., Eisenberg, A.J., Budowle, B., Taha A., Miller, R.W., McGuckian, A.B., Conover- Sikorsky, J., Crouse, C.A. Assessing a novel room temperature DNA storage medium for forensic biological samples For. Sci. Int. - Gen. in press.

10 High Volume Workflow Thompson, et. al. - Orange County Crime Lab - DNAstable LD Body fluids and Standards Evidence Exam (samples start in tubes) EZ1 DNA Extraction (extracts in tubes) Universal qpcr set-up qpcr Universal amp Set-up Amp Corbett/ Qiagility CE setup 3130 Genetic Analyzer Trace Tecan DNA Extraction (extracts in plates) Universal qpcr set-up qpcr Universal amp Set-up Amp Corbett/ Qiagility CE setup 3130 Genetic Analyzer Add DNAstable LD to samples not consumed Incorporation of DNAstable LD into a High Volume DNA Analysis Line, by Jennifer Jarrett, Forensic Scientist II and Elizabeth Thompson, DNA Lab Director, Orange Cty Crime Lab, CA. Workshop on Forensic DNA Analysis and Storage at the 2011 AAFS Meeting.

11 Returning extracts to source WSP is required to ship back remaining DNA extracts with evidence to submitting agencies Evaluated DNAstable LD internally prior to adoption 29 samples analyzed (SM-preserved, stored, recovered and then reanalyzed between 5 labs) Before & after quantification performed Excellent recovery (within variation expected for Quantifiler) Before & after genotypes compared All concurrent with no contamination Extracts now shipped in DNAstable starting 2010 Return of DNA extracts to submitting agencies, by Dr Gary Shutler, DNA Tech Lead/Manager, Forensic Lab Services Bureau, WA State Patrol, Seattle, WA, Workshop on Forensic DNA Analysis and Storage, 2011 AAFS Meeting.

12 CONFIDENTIAL

13 Forensic-based publications on DNAstable /QIAsafe DNA studies Peer-reviewed articles ( ) Lee, S.B., Clabaugh, K.C., Silva, B., Odigie, K.O., Fourney, R.M., Stevens, J., Carmody, G.R., Coble, M.D., Loreille, O., Scheible, M., Parsons, T.J., Pozder, A., Eisenberg, A.J., Budowle, B., Taha A., Miller, R.W., McGuckian, A.B., Conover-Sikorsky, J., Crouse, C.A. Assessing a novel room temperature DNA storage medium for forensic biological samples For. Sci. Int. - Gen. in press. Lee S.B., Crouse, C.A. and Kline M.C. Optimizing Storage and Handling of DNA extracts; Forensic Sci. Rev., 2010, 22(2): Frippiat, C., Zorbo, S., Leonard, D., Marcotte, A., Chaput, M., Aelbrecht, A., Noel, F. Evaluation of novel forensic DNA storage methodologies, Forensic Sci. Int. Genetics, Budowle, B., van Daal, A. Extracting evidence from forensic DNA analyses: future molecular biology directions, Biotechniques, 2009, 46 (5), Poster Presentations ( ) Breeana Baker, Clarissa Trogdon and Steven Lee, Analysis of (N-4)bp STR Repeat Slippage on Low Quantity DNA Samples with PCRboost, 63rd AAFS Meeting, Chicago, IL, March 22-25, Kevin Kramer, Brent Myers, Heather Harrah-Lea, and Pamela Staton, Comparison of Room Temperature Forensic DNA Extract Sample Preservation Methods. 63rd AAFS Meeting, Chicago, IL, March 22-25, Erica DiNaro, Clarissa Trogdon and Steven B. Lee, Optimizing DNA Storage at Room Temperature: Teflon vs. Polypropylene Tubes With and Without Polymers. 63 rd AAFS Meeting Gregory M. Hadinoto. Evaluation of the Biomatrica SampleMatrix room temperature DNA storage system: 12-month and 45-month data. Poster #60, 21st International Symposium on Human Identification, San Antonio, TX, Oct 11-14, More information can be found at:

14 Using, Adopting or Testing Palm Beach County Sheriff s Office, FL Washington State Patrol, WA Orange County Crime Lab, CA Hennepin County Sheriff s Office, MN FBI, Quantico, VA LA County Sheriff s Office, CA New York Office of the Chief Medical Examiner Harris County Institute of Forensic Science, TX SWIFS, TX San Diego Sheriff s Department, CA San Bernardino Sheriff s Department, CA Cal-DOJ San Jose State University Prof. Steven Lee Eastern Washington University Prof. Peter Bilous Boise State University Prof Greg Hampikian UNT HSC Prof Bruce Budowle..etc. Evaluation studies as presented at a 2011 AAFS Workshop available for download

15 Conclusions Long-term DNA storage with high quality, better recovery and a cost-effective solution offered Cost effective alternative to cold freezers Safety & security in event of natural disaster Applicable in manual or automated casework protocols Multiple storage vessels: tubes, plates, custom Multiple reagent formats: dry or liquid Extensive testing and validation already conducted Articles, posters, talks, etc Offer validation support (kit) for resource/time-crunched forensic labs Contact us for details (contact@biomatrica.com) Sample management software to capture, track and manage all stages of collection, shipping, processing and storage. SampleWare

16 More information (858) (866)