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Stabbing simulations and DNA transfer.

FSI Genetics, 2016

Study Design

Addressed Question

study of the transfer of DNA from handler and closely connected individuals to a knife handle

Activity Context

Stabbing

Category

Primary Deposit

Specifications

Individual Characteristics

Variables of Interest

individualsprevious contactstime of the day

Stringency of Control

Controlled

Number of Individuals

4

Replicates per Individual and Condition

16

Nucleic Acid

DNA

Bodily Origin

skin (hands)

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

Plastic

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

Negative (Confirmed)

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