DNA transfer by examination tools - A risk for forensic casework?
FSI Genetics, 2015
Authors
Journal
FSI Genetics
Study Design
Addressed Question
occurrence and level of DNA transfer via high risk vectors under different contact scenarios and its impact on target sample DNA profiling
Activity Context
Category
Specifications
Variables of Interest
Stringency of Control
Number of Individuals
2
Replicates per Individual and Condition
3-6
Nucleic Acid
Bodily Origin
Depositor & Contact
Depositor Characteristics
one male, one female
Criteria for Shedder Status
N/A
Previous Activities
N/A
Contact Scenario
deposit on primary and secondary substrate - drying time (12-24 h) - contact primary substrate + vector - contact secondary substrate + vector - sampling
Primary Substrate
Primary Substrate Type
A4 piece of cotton fabric drill, glass slide
Primary Substrate Material
Deposit
25 µl venous blood, vigorous rubbing 30s on cotton, pressure 30s on glass
Delay
N/A
Secondary Substrate
Secondary Substrate Type
metal tools (scissors, forceps), nitrile gloves
Secondary Substrate Material
Secondary Substrate Contact
light (medium pressure touch/cut 2-3s), heavy (8x medium pressure touch/cut 2-3s)
Further Transfer
cotton, glass light or heavy contact
Sampling
Background DNA on Sampled Surface
Sampling Time
direct
Persistence
N/A
Sampling Method
cotton swabs (double swabbing) for vectors and glass slides, direct extraction from cotton substrate
Sampling Area
1.5x1.5 cm^2 contact areas
Laboratory Analysis
Extraction
DNA IQ extraction system (Promega)
DNA Quantification
Quantifiler (Life Technologies), ABIPRISM7500 (Life Technologies), SDS software
Input for Profiling
0.5 ng or 15 µl of DNA template
Profiling
PowerPlex 21 System (Promega), ABIPRISM 3500xL Genetic Analyzer, GeneMapper ID-X software, threshold: 175 rfu
Reference Samples
taken from all participants
Profile Interpretation and Mixture Analysis
comparison to reference profiles, interpretation of unique alleles only, peak heights (rfu) used for the determination of major/minor contributors
RNA Data Interpretation
N/A
Results
DNA Quantity
n.s. (Supp.: 0.000-0.016 ng/µl)
Profile Quality
deposits: full profiles from blood, high variability from touch deposits (35-100%), unique unknown alleles: max 32 in touch, max 5 in blood samples; secondary profiles: full profiles; transferred profiles: partial profiles (0-30%)
Parameter Used for Comparison
% of unique alleles transferred, corresponding average unique peak height
Summary of Results
similar patterns observed as in experiment with DNA-free backgrounds but significantly fewer unique alleles detected from primary deposit when secondary substrate not DNA free; the transferred alleles were mostly minor contributors or absent (even for dried blood transferred to touch DNA); detectability of secondary profile (background DNA) not affected by transfer scenarios
Raised Questions
potential for intra-exhibit transfer; loss of DNA in the exhibit examination process
Cautionary Remarks
reduced comparability for % transferred alleles from touch DNA due to high variation of initial deposit (between blood and touch as well as among touch DNA samples), contact scenario rather unrealistic (no prevention of contamination); non-donor alleles observed in blood positive controls; mixture interpretation approach not performed according to casework procedure; % observed unique allelic peaks and average peak heights both prone to stochastic effects