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DNA transfer during laundering may yield complete genetic profiles.

FSI Genetics, 2016

Study Design

Addressed Question

transfer of epithelial cells from vaginal secretions during washing

Activity Context

Sexual ContactWashing Machine

Category

RecoveryTransfer Scenario

Specifications

Bodily OriginSampling

Variables of Interest

donorspre- vs. Postmenopausal stage

Stringency of Control

Close to Realistic

Number of Individuals

2

Replicates per Individual and Condition

1 (5)

Nucleic Acid

DNA

Bodily Origin

vaginal secretions

Depositor & Contact

Depositor Characteristics

one pre-menopausal, one post-menopausal woman, no menstruation, no sexual contact within the last 5 days

Criteria for Shedder Status

N/A

Previous Activities

N/A

Contact Scenario

wearing of 5 different pieces of underwear for 1 day each - washing with 5 pieces of clean underwear and 1 bedsheet - sampling

Primary Substrate

Primary Substrate Type

100 % cotton fabric underwear

Primary Substrate Material

Cotton

Deposit

wearing for 1 day

Delay

N/A

Secondary Substrate

Secondary Substrate Type

cotton fabric underwear

Secondary Substrate Material

Cotton

Secondary Substrate Contact

washing: 30°C + cold water rinse + phosphate free detergent, medium heat drying 60 min

Further Transfer

N/A

Sampling

Background DNA on Sampled Surface

Negative (Assumed)

Sampling Time

direct

Persistence

N/A

Sampling Method

swabbing and cutting

Sampling Area

swabbing 40 cm^2, cuttings 6.25 cm^2, 6-7 different areas per piece of underwear and sampling method

Laboratory Analysis

Extraction

differential extraction DNA IQ magnetic beads system (Promega)

DNA Quantification

Quantifiler Duo DNA Quantification kit (AB), 7500 Real-Time PCR

Input for Profiling

set volume 15 µl (min 0.008 ng/µl for epithelia fraction, 0.001 ng/µl for spermatozoa fraction)

Profiling

AmpFlSTR Identifiler Plus PCR Amplification kit 28 cycles, ABI PRISM 3130xI Genetic Analyzer, GeneMapper ID software v.3.2, AT: 50rfu, ST: 200 rfu

Reference Samples

taken from wearers

Profile Interpretation and Mixture Analysis

determination of number of alleles from reference profiles

RNA Data Interpretation

N/A

Results

DNA Quantity

<amplification threshold for swabbings, 0.2-3.2 ng for cuttings

Profile Quality

partial to complete profiles from samples with highest amplification results

Parameter Used for Comparison

DNA quantity, profile completeness

Summary of Results

higher amounts of DNA from pre-menopausal woman; partial to complete genetic profiles obtainable

Raised Questions

N/A

Cautionary Remarks

sample size too low for differentiation between pre- and post-menopausal stage (n=1); quality of DNA profiles (peak height contributions, unknown alleles n.a.)