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
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 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 profile, determination of % of total alleles observed and total corresponding peak heights
RNA Data Interpretation
N/A
Results
DNA Quantity
n.s. (Supp.: 0.000-0.008 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; transfer: partial profiles (10-95%) transferred from dried blood, no or partial profiles (0-20%) transferred from touch;
Parameter Used for Comparison
% transferred alleles, average peak height (RFU)
Summary of Results
Transfer rates dependent on transfer scenario, general trends: dried blood >> touch DNA, gloves > scissors > forceps, heavy/heavy > heavy/light, light/light contact, primary substrate: cotton > glass for blood, glass > cotton for touch DNA; acceptor substrate: cotton > glass
Raised Questions
minimization of inter-exhibit transfer by following cleaning protocols, comparison to findings from Goray et al (2010): found that plastic facilitates transfer of blood, whereas in this case cotton was better -> glass vs plastic, flaking of blood?
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); only secondary substrate sampled, it can thus not be differentiated between retention by primary substrate or retention by vector (cf. Szkuta et al. (2015): Residual DNA on examination tools following use); contact scenario rather unrealistic (no prevention of contamination); non-donor alleles observed in blood positive controls; % observed unique allelic peaks and average peak heights both prone to stochastic effects