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DNA transfer by examination tools - A risk for forensic casework?

FSI Genetics, 2015

Study Design

Addressed Question

occurrence and level of DNA transfer via high risk vectors under different contact scenarios

Activity Context

Professional

Category

Transfer Scenario

Specifications

Bodily OriginContactSurfaceTransfer via Vector

Variables of Interest

contact surfacestransfer vectorcontact scenariobiological substrate

Stringency of Control

Controlled

Number of Individuals

2

Replicates per Individual and Condition

3-6

Nucleic Acid

DNA

Bodily Origin

bloodskin (hands)

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

CottonGlass

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

MetalNitrile

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

Negative (Confirmed)

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