Stabbing simulations and DNA transfer.
FSI Genetics, 2016
Authors
Journal
FSI Genetics
Study Design
Addressed Question
study of the transfer of DNA from handler and closely connected individuals to a knife handle
Activity Context
Category
Specifications
Variables of Interest
Stringency of Control
Number of Individuals
4
Replicates per Individual and Condition
16
Nucleic Acid
Bodily Origin
Depositor & Contact
Depositor Characteristics
two males, two females, two left-handed, two-right-handed
Criteria for Shedder Status
N/A
Previous Activities
handwashing in the morning, normal activities
Contact Scenario
handwashing 8am - delay - stabbing 11am - delay - stabbing 4pm
Primary Substrate
Primary Substrate Type
plastic knife handle
Primary Substrate Material
Deposit
stabbing simulation
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
Swabbing (wet + dry)
Sampling Area
knife handles
Laboratory Analysis
Extraction
QIAshredder and QIAamp kit, Microcon 30 spin columns
DNA Quantification
Investigator Quantiplex Hyres (Qiagen)
Input for Profiling
a.p.m.i.: 1 ng in 10 µl
Profiling
NGM Select 30 cycles in triplicates, 3130 Genetic Analyzer ABI, GeneMapper IDX software, AT: 30 rfu
Reference Samples
taken from all depositor + colleagues
Profile Interpretation and Mixture Analysis
evaluation using STRmix v.2.3.05: number of contributors based on peak heights and peak height balance, determination of mixture proportions (major: >70%), assignment of major and minor contributors for weights >0.7, profile fully represented, when combinations has a weight >0.7 for all loci, and partial but of value when weight >0.7 for at least 6 loci; major and minor DNA profiles compared to reference profiles and colleagues met during the day
RNA Data Interpretation
N/A
Results
DNA Quantity
50 - 485 pg
Profile Quality
peak height: 50-600 rfu, mostly mixed profiles with stabber as full or partial major profiles, profile rarely unsuitable for comparison (5%)
Parameter Used for Comparison
(DNA yield), profile completeness, relative contributor quantities
Summary of Results
high intra and inter-person variability regarding quantification results; no clear correlation between DNA concentrations and quality of obtained profiles, especially for low DNA concentrations high variability -> relative quantity and profile quality as a better descriptor; DNA of stabber present as major profile in 83 %, 3% as single profile, otherwise mixtures from 2,3 and 4 contributors; no full non-matching profile observed; profile unsuitable for comparison in 5 % (>3 contributors); binary approach would have led to false exclusions in some cases, continuous approach more useful; stochastic effects can lead to the appearance of stabber as minor contributor in some alleles and major in others; stabber's colleagues' profiles not observed; no significant difference between experiments performed at 11 am and 4 pm
Raised Questions
What are extrinsic DNA characteristics (concentration, quality or relative quantity of profile)? Investigation of background DNA on knife handles; source of background DNA profiles on hands (household members)? Processing knife as if it was casework (e.g. fingerprint detection)
Cautionary Remarks
intensity and duration of stabbing scenario not specified; maximum chance conditions due to DNA-free knifes; individual characteristics or previous activities not further considered; sources other than colleagues not taken into consideration for minor contributors