Comparison of DNA Stability Using Various Sample Collection Devices - - PowerPoint PPT Presentation

Criminal Investigation Laboratory, Oita Prefectural Police, Japan Seiki Nakao

Comparison of DNA Stability Using Various Sample Collection Devices for forensic DNA analysis

Today’s Presentation

• 1. Introduction
• 2. Materials and Methods
• 3. Results
• 4. Summary and Further works

• 1. Introduction
• 2. Materials and Methods
• 3. Results
• 4. Summary and Further works

Today’s Presentation

DNA collection devices

Medical Swab Forensic Swab Adhesive Tape All samples blood samples touch samples

DNA collection devices

Divisible 4N6FLOQSwab

Divisible 4N6FLOQSwab Medical Swab Forensic Swab Adhesive Tape

Comparison

l DNA recovery l DNA stability

Objectives

Today’s Presentation

• 1. Introduction
• 2. Materials and Methods
• 3. Results
• 4. Summary and Further works

Sample Preparation and Collection

Stored RT for 1 week, 2 and 4 weeks Swabbing by wetted swabs Tape lifting (15 times)

Dried O/N

Saliva Blood 20 μl

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 3130xl 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

1 week 2 week 4 week

decayed decayed molded

Medical Swab Forensic Swab Divisible 4N6FLOQSwab

Swabs after sampling blood

1 week 2 weeks 4 weeks

Tapes after sampling blood

molded molded

1 week 2 week 4 week Medical Swab Forensic Swab Divisible 4N6FLOQSwab

Swabs after sampling saliva

1 week 2 weeks 4 weeks

Tapes after sampling saliva

DNA recovery and stability of blood sample

(0.50) 0.00 0.50 1.00 1.50 2.00

50 100 150 200 250 300 350 400

• 1

1 1 2 2 50 100 150 200 250 300 350 400 Ctrl-血 液-BTA

• 1

1 1 2 2 50 100 150 200 250 300 350 400

• 1

1 1 2 2 50 100 150 200 250 300 350 400

• 1

1 1 2 2

50 100 150 200 250 300 350 400

Ctrl-血 液-PF

Total DNA(ng) Degradation Index

• 1

1 1 2 2

Medical swab Divisible 4N6FLOQ Swab Adhesive tape Liquid- Blood- Ctrl(PF) Forensic Swab Liquid- Blood- Ctrl(BTA)

1 2 4 1 2 4 1 2 4 1 2 4 (week(s))

• 0.5

0.0 0.5 1.0 1.5 2.0 2.5 20 40 60 80

• 0 .

20 40 60 80

• 0.5

0.0 0.5 1.0 1.5 2.0 2.5 20 40 60 80 Ctrl-唾液-PFBTA

• 0 .

20 40 60 80

Ctrl-唾 液-PF

Total DNA(ng) Degradation Index

• 0 .

20 40 60 80

(0.50) 0.00 0.50 1.00 1.50 2.00 2.50

DNA recovery and stability of saliva sample

Medical swab Divisible 4N6FLOQ Swab Adhesive tape Liquid- Blood- Ctrl(PF) Forensic Swab Liquid- Blood- Ctrl(BTA)

1 2 4 1 2 4 1 2 4 1 2 4 (week(s))

Electropherogram of blood sample（1）

1 week

Medical Swab Forensic Swab Divisible 4N6FLOQSwab Adhesive Tape

DI=0.60 DI=1.43 DI=0.63 DI=0.60

2 weeks

Medical Swab Forensic Swab Divisible 4N6FLOQSwab Adhesive Tape

Electropherogram of blood sample（2）

DI=1.03 DI=1.50 DI=0.63 DI=0.70

Electropherogram of blood sample（3）

4 weeks

Medical Swab Forensic Swab Divisible 4N6FLOQSwab Adhesive Tape

DI=0.80 DI=1.56 DI=0.66 DI=0.67

Electropherogram of saliva sample（1）

1 week

Medical Swab Forensic Swab Divisible 4N6FLOQSwab

DI=0.63 DI=1.33 DI=0.60

Electropherogram of saliva sample（2）

2 weeks

Medical Swab Forensic Swab Divisible 4N6FLOQSwab

DI=0.73 DI=1.53 DI=0.67

Electropherogram of saliva sample（3）

4 weeks

Medical Swab Forensic Swab Divisible 4N6FLOQSwab

DI=0.70 DI=1.73 DI=0.75

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

• n 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.