Comparison of DNA Stability Using Various Sample Collection Devices for forensic DNA analysis Criminal Investigation Laboratory, Oita Prefectural Police, Japan Seiki Nakao
Today’s Presentation 1. Introduction 2. Materials and Methods 3. Results 4. Summary and Further works
Today’s Presentation 1. Introduction 2. Materials and Methods 3. Results 4. Summary and Further works
DNA collection devices All samples Medical Swab Forensic Swab blood samples Adhesive Tape touch samples
DNA collection devices Divisible 4N6FLOQSwab
Objectives Medical Swab Forensic Swab Adhesive Tape l DNA recovery Comparison l DNA stability Divisible 4N6FLOQSwab
Today’s Presentation 1. Introduction 2. Materials and Methods 3. Results 4. Summary and Further works
Sample Preparation and Collection Saliva Blood 20 μl Dried O/N Swabbing by Tape lifting wetted swabs (15 times) Stored RT for 1 week, 2 and 4 weeks
Extraction and Quantification Extraction Kit ( Swab) PrepFiler Express Forensic DNA Extraction Kit (Applied Biosystems) (Adhesive Tape) PrepFiler Express BTA Forensic DNA Extraction Kit (Applied Biosystems) Quantification Quantifiler Trio DNA Quantification Kit (Applied Biosystems)
DNA Amplification and Profiling DNA Amplification Identifiler Plus Kit (Applied Biosystems) Electrophoresis Applied Biosystems 3130 xl Genetic Analyzer (Applied Biosystems) Analysis GeneMapper ID-X (Applied Biosystems)
Today’s Presentation 1. Introduction 2. Materials and Methods 3. Results 4. Summary and Further works
Swabs after sampling blood 1 week 2 week 4 week decayed decayed Medical Swab molded Forensic Swab Divisible 4N6FLOQSwab
Tapes after sampling blood 1 week 2 weeks 4 weeks
Swabs after sampling saliva 1 week 2 week 4 week molded Medical Swab molded Forensic Swab Divisible 4N6FLOQSwab
Tapes after sampling saliva 1 week 2 weeks 4 weeks
DNA recovery and stability of blood sample 2.00 400 400 400 2 400 2 400 2 2 2 4 0 350 350 350 350 350 3 5 0 1.50 2 2 2 2 2 300 300 300 300 Degradation Index 300 3 0 Total DNA(ng) 250 250 250 250 250 1.00 1 1 2 5 0 1 1 1 200 200 200 200 200 2 0 0.50 1 1 1 1 1 150 150 150 150 150 1 5 0 100 100 100 100 100 1 0 0.00 0 0 0 0 0 50 50 50 50 50 5 0 (0.50) 0 0 0 -1 -1 -1 0 0 -1 -1 0 Ctrl- 血 1 2 4 1 2 4 1 2 4 1 2 4 (week(s)) Ctrl- 血 液 -BTA 液 -PF Liquid- Medical Forensic Swab Divisible Liquid- Adhesive Blood- swab 4N6FLOQ Blood- tape Ctrl(PF) Swab Ctrl(BTA)
DNA recovery and stability of saliva sample 80 2.50 80 80 80 2.5 80 2.5 2 . 5 2 . 5 8 0 0 . 2 . 5 2.00 2.0 2.0 2 . 0 2 . 0 2 . 0 60 60 60 60 60 Degradation Index 6 0 0 . Total DNA(ng) 1.50 1.5 1.5 1 . 5 1 . 5 1 . 5 40 1.00 40 40 40 1.0 40 1.0 1 . 0 1 . 0 4 0 0 . 1 . 0 0.50 0.5 0.5 0 . 5 0 . 5 0 . 5 20 20 20 20 20 2 0 . 0 0.00 0.0 0.0 0 . 0 0 . 0 0 . 0 0 (0.50) 0 0 0 -0.5 0 -0.5 0 . - 5 - 0 . 5 0 . 0 0 . - 5 1 2 4 1 2 4 1 2 4 1 2 4 (week(s)) Ctrl- 唾 Ctrl- 唾液 -PFBTA 液 -PF Liquid- Medical Forensic Swab Divisible Liquid- Adhesive Blood- swab 4N6FLOQ Blood- tape Ctrl(PF) Swab Ctrl(BTA)
Electropherogram of blood sample （ 1 ） 1 week DI=0.60 Medical Swab DI=1.43 Forensic Swab DI=0.63 Divisible 4N6FLOQSwab DI=0.60 Adhesive Tape
Electropherogram of blood sample （ 2 ） 2 weeks DI=1.03 Medical Swab DI=1.50 Forensic Swab DI=0.63 Divisible 4N6FLOQSwab DI=0.70 Adhesive Tape
Electropherogram of blood sample （ 3 ） 4 weeks DI=0.80 Medical Swab DI=1.56 Forensic Swab DI=0.66 Divisible 4N6FLOQSwab DI=0.67 Adhesive Tape
Electropherogram of saliva sample （ 1 ） 1 week DI=0.63 Medical Swab DI=1.33 Forensic Swab Divisible DI=0.60 4N6FLOQSwab
Electropherogram of saliva sample （ 2 ） 2 weeks DI=0.73 Medical Swab DI=1.53 Forensic Swab Divisible DI=0.67 4N6FLOQSwab
Electropherogram of saliva sample （ 3 ） 4 weeks DI=0.70 Medical Swab DI=1.73 Forensic Swab Divisible DI=0.75 4N6FLOQSwab
Summary l The Divisible 4N6FLOQSwab was the most effective upon the DNA recovery and stability from saliva and blood sample on the glass slide. l Forensic swab was effective on the DNA recovery from blood sample but not from saliva sample and a little lower on its satabilty from both samples. l Medical swab was easy to be molded and decayed and DNA yield from it was low. l Adhesive tape was effective on the DNA recovery and stability from blood sample but not on saliva sample at all.
Further works l Influence of storage temperature on samples. l Collecting touch type sample.
Acknowledgements Criminal Investigation Laboratory, Oita Prefectural Police Eiji Nishi Yoshito Tomisaka Takaaki Ono Nao Mori Kohei Tomonari Ami Ikehara Ayane Sonoda Life Sciences Solution Group, Thermo Fisher Scientific Takekiyo Honma Kyo Azuma
THANK YOU! Speaker was provided travel and hotel support by Thermo Fisher Scientific for this presentation, but no remuneration. When used for purposes other than Human Identification or Paternity Testing the instruments and software modules cited are for Research Use Only. Not for use in diagnostic procedures. Thermo Fisher Scientific and its affiliates are not endorsing, recommending, or promoting any use or application of Thermo Fisher Scientific products presented by third parties during this seminar. Information and materials presented or provided by third parties are provided as-is and without warranty of any kind, including regarding intellectual property rights and reported results. Parties presenting images, text and material represent they have the rights.
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