Typeability of DNA in touch traces deposited on paper and optical data discs.
Advances in Clinical and Experimental Medicine, 2015
Authors
- MSMałgorzata Skawrońska
- WPWitold Pepiński
- ISIreneusz Sołtyszewski
- MSMichał Szeremeta
- ANAnna Niemcunowicz-Janica
Journal
Advances in Clinical and Experimental Medicine
Study Design
Addressed Question
DNA typing from fingerprints using standard and LCN conditions
Activity Context
Category
Specifications
Variables of Interest
Stringency of Control
Number of Individuals
60
Replicates per Individual and Condition
3
Nucleic Acid
Bodily Origin
Depositor & Contact
Depositor Characteristics
30 males, 30 females
Criteria for Shedder Status
N/A
Previous Activities
handwashing, 2h of office activities and touching face/hair
Contact Scenario
handwashing - delay - finger deposit - sampling
Primary Substrate
Primary Substrate Type
office paper, optical data discs (CDs and DVDs)
Primary Substrate Material
Deposit
pressing/rubbing three fingers
Delay
N/A
Secondary Substrate
Secondary Substrate Type
N/A
Secondary Substrate Material
N/A
Secondary Substrate Contact
N/A
Further Transfer
N/A
Sampling
Background DNA on Sampled Surface
Sampling Time
direct
Persistence
N/A
Sampling Method
dry swabbing from data discs, direct extraction from paper
Sampling Area
deposit area from optical discs, 2x2cm^2 cuttings from paper
Laboratory Analysis
Extraction
QIAamp DNA Mini kit modified for epithelial cells
DNA Quantification
Quantifiler Human DNA Quantification kit, 7500 real time PCR
Input for Profiling
N/A
Profiling
AmpFlSTR NGM amplification kit at standard 30 and LCN 34 cycles (in triplicates), 3130 Genetic Analyzer, GeneMapper ID-X v1.1.1, threshold: 50/200 rfu; alleles scored when >5% of the most prominent peak at the locus
Reference Samples
buccal swabs taken from all depositors
Profile Interpretation and Mixture Analysis
An allele was scored when its peak height was > 5% of the peak height of the most prominent allele at a given locus; generation of consensus profiles from two replicate reactions; assignment of a profile, when at least 5 loci typeable, comparison to reference profiles and determination of "false" alleles and drop-outs
RNA Data Interpretation
N/A
Results
DNA Quantity
0-1.2 ng from CDs and 0-0.8 ng from paper
Profile Quality
mostly partial, some full and some null profiles
Parameter Used for Comparison
DNA profile detectability (full, partial, null (<5 alleles), DNA yield, rfu range
Summary of Results
large interindividual differences; no significant differences between paper and data discs, no significant difference between male and female depositors; 30 vs. 34 cycle protocol: increased success rate (about 10%), increased average rfu of peaks, increased detection of additional alleles in non-stutter positions
Raised Questions
N/A
Cautionary Remarks
origin of additional alleles not questioned (treated as artefacts but could also be from secondary transfer); consensus approach described in method section but not clear whether it was applied in the results section (i.e. was a reduction of drop-in by the consensus approach achieved?); increased deposit chance due to repeated touching of hair and face in the 2h prior to deposit; no statistical evaluation of results