Forensic Science International: Genetics 8 (2014) 219–225

Contents lists available at ScienceDirect

Forensic Science International: Genetics

journal homepage: www.elsevier.com/locate/fsig

Persistence of DNA deposited by the original user on objects after

subsequent use by a second person
Roland A.H. van Oorschot a,*, Genna Glavich a,b, R. John Mitchell b
a
Victoria Police Forensic Services Department, 31 Forensic Drive, Macleod, Melbourne, Victoria 3085, Australia
b
Department of Genetics, School of Molecular Sciences, La Trobe University, Melbourne, Victoria 3086, Australia

A R T I C L E I N F O A B S T R A C T

Article history: There is a paucity of data regarding the persistence of DNA from prior user of an object after, its use by
Received 3 April 2013 another person. To acquire a greater understanding of persistence we performed controlled, experiments
Received in revised form 29 August 2013 encompassing over 179 objects that had only been used by one individual for an extended, period before
Accepted 9 October 2013
used by a 2nd person for various but known duration. Our findings show that the profile, percentage
contribution of the 1st user relative to the 2nd user of an object declines in a linear manner, over time.
Keywords: The retrieval of the profile of the initial user of the object is dependent on the type of, substrate and use of
DNA
the object. When considering a hard non-porous object the 1st user’s profile, percentage contribution
Persistence
drops 50% immediately upon use by a 2nd person and drops to 15% after, 90 min. When considering a
Transfer
Touch soft porous object the 1st wearer’s profile contribution remains, higher than that of the 2nd wearer
Forensic during the first 10 h of wear by the 2nd wearer and still, accounts for 12% after 96 h. This substrate
associated difference was also observed in an, assessment of a wide range of personal objects used by
2nd users for different durations. Particular, areas of certain objects were more likely to retain a greater
proportion of the 1st user’s DNA than other, areas. Alleles of unknown source were present on the
majority of objects but rarely exceeded 10% of, the total profile. Greater knowledge of persistence will
inform investigators regarding the likelihood of, detecting a profile of a particular individual based on the
type of object and its history, and assist with, identifying the best areas of an object to target for DNA
sampling.
ß 2013 Elsevier Ireland Ltd. All rights reserved.

1. Introduction weapon, tool equipment, keyboard, phone) or come into contact

Crime investigators regularly encounter exhibits that have been
used/worn for an extended period of time by one person (usually
the owner) then subsequently used/worn by a second person. The
time that the second person uses/wears the object could vary
enormously, from a brief moment to an extended length of time.
The DNA profile of the last user/wearer of an item is often
retrievable from a touched item [1,2]. Whilst it is frequently the
last user that investigators are interested in, sometimes it is the
previous user that is the focus of attention. Examples of this can
include: identifying the owner of a stolen item; identifying a prior
user of a weapon that may have been shared; seeking confirmation
of scenarios relating to criminal activity involving a specific
individual who may have used a specific item (i.e. clothing,
* Corresponding author at: Office of the Chief Forensic Scientist, Forensic Services
Centre, 31 Forensic Drive, Macleod 3085, Victoria, Australia. Tel.: +61 3 9450 3528;
fax: +61 3 9450 3601.
E-mail addresses: roland.vanoorschot@police.vic.gov.au
(Roland A.H. van Oorschot), john.mitchell@latrobe.edu.au (R.J. Mitchell).
with a particular surface (i.e. door, window, furniture) which is
known to have been used/touched by others since. When
evaluating the chance of obtaining an individual’s profile from a
particular exhibit with a given history, it is valuable to understand
the persistence of DNA on such exhibits. This knowledge is relevant
for sampling prioritisation and targeting, as well as interpretation
of profiles in case investigations. In this context an important
question is: to what extent can the DNA profile of the former user/
wearer of an object still be detected after use by a second person?
Knowledge of persistence in given circumstances may also provide
an indication of the time that has lapsed between events.
It is well accepted that biological samples collected from certain
surfaces often provide a mixture of profiles [1,3–6]. This is
particularly true when the target samples are of a trace nature. In
such situations the underlying DNA (often also present in trace
quantities) constitutes a higher proportion of the DNA sample
collected. Examples include assaults where the victims’ clothing
has been touched, steering wheels of stolen cars, and personal
objects, such as wallets, handbags, watches, jewellery that have
been stolen, and weapons owned by an individual that have
temporarily been used by someone else.

1872-4973/$ – see front matter ß 2013 Elsevier Ireland Ltd. All rights reserved.
http://dx.doi.org/10.1016/j.fsigen.2013.10.005
220 R.A.H. van Oorschot et al. / Forensic Science International: Genetics 8 (2014) 219–225
There have been very few studies examining persistence of
biological samples [4,7] and even fewer that focussed on the
persistence of DNA left by touch/skin cells, especially after the
object has been used/worn by a second person. One such study
showed that the majority of profiles recovered from wallets stolen
in a simulated robbery were mixtures, with the ‘robber’s profile
comprising the major component of the mixture or was the single
source in 40% of profiles [8]. The authors concluded that such
knowledge of trace evidence characteristics of DNA will aid in its
interpretation and presentation in criminal trials.
Transfer of DNA from touched objects is dependent on the
substrate on which the biological material resides, the substrate
the deposit area comes into contact with, the freshness of the
deposit and the manner of contact [9]. There appear to be other
factors, however, also influencing transfer [10]. The value of an
awareness of transfer rates given particular situations to help
judge the relative probability of alternative multi-step transfer
events has also been illustrated [11].
Here we aim to acquire a greater understanding of the
persistence of DNA deposited by the initial user on objects after
various durations of use of the item by a second person. Three
separate tests were conducted: Test A relating to a handled object
that had a non-porous, hard, flat surface (represented by pens and
pen lids); Test B relating to a worn object that was porous
(represented by bracelets made of elastic fabric); and Test C, where
unlike Tests A and B where all the starting objects were the same,
we examine a wide range of everyday personal objects of different
substrates, shapes, sizes and uses. All Test A and B replicate objects
had similar determinable amounts of DNA deposited on them by
an initial known single individual, while the objects used in Test C
had varying unknown amounts of DNA, each from a different initial
depositor. All objects from Tests A, B and C were given to a wide
range of second users who used them for different periods of time.
2. Materials and methods
2.1. Objects and activity recordings
2.1.1. Test A
Fifty-four new pens (Classic Fine blue ink, Bic Australia)
hexagon shaped hard smooth plastic with a cone shaped hard
smooth plastic lid taken directly from a new box were cleaned with
0.05% hypochlorite and 70% ethanol. Five pens, with their lids, were
retained as negative controls and all other pens were placed
together in a single large envelope. To ensure that, as much as
possible, an approximately equal amount of DNA, from a known
single donor source, was placed on each pen they were handled as
follows: Five pens were randomly taken from the envelop at a time,
rubbed between two hands for 30 s, the lid of each removed and
replaced, and placed on clean sheet of blotting paper. After all pens
had been treated in this manner they were returned to the
envelope. This procedure was repeated (including the removal and
replacement of lids) on each of the following three days by the
same person, but instead of rubbing the pens for 30 s they were
rubbed for 60 s. In total each pen had been rubbed vigorously for a
total of 210 s and the lids removed and replaced four times. Five of
these pens were randomly removed from the large envelope using
clean forceps and placed in a smaller envelope as deposit controls.
Using clean forceps each of the remaining pens was removed from
the large envelope and placed into a separate small envelop. 44 of
these were issued to second users (each a different individual).
The second users were provided with an information sheet, an
envelope with a single used pen, an activity data recording sheet
and a new, small, empty envelope into which the pen was to be
placed upon completion of use. The activities to which the pen was
exposed were recorded on the data sheet and these details
included; the date and time of each change of activity; the type of
material with which it was in contact during the activity (including
various skin locations and non skin substrate types); description of
substrate type; the type of activity the object was involved in and
the manner of contact, and the duration of the activity. The second
users were requested to use the pen as they normally would for
writing and record contact information. Buccal samples were taken
from all 1st and 2nd users from which their profiles were
generated.
2.1.2. Test B
A stretch of flat white nylon/polyester blend braided 3 mm
wide elastic (purchased from a local chain store) was cut into
pieces of 24 cm lengths using cleaned scissors. The ends of each
piece were knotted and the overhanging elastic cut from the knots
to create 88 elastic band bracelets of 19 to 22 cm. All bracelets were
laid flat on blotting paper and both sides exposed to UV light for
12 h respectively. Six bracelets were put aside as negative controls.
To ensure that an approximately equal amount of DNA, from a
known single donor source, was deposited on each bracelet they
were handled as follows: A single individual wore 80 bands (40 on
each arm) between the hand and the elbow for a total of 34 h
during 6 sessions extending over 5 days. Prior to removing the
bracelets after each of these 6 sessions the wearer rubbed his left
hand over the bracelets on the right arm for 30 s and the same with
his right hand over the bracelets on his left arm. Each time the
bracelets were removed from the arm the wearer placed them
together in a single large envelope then rummaged through the
envelope and randomly handled bunches of bracelets for 1 min
with his right hand and 1 min with his left hand. The same person
then randomly removed 4 bracelets from the envelope, rubbed
them between his hands for 30 s and placed them in a small
envelope. This procedure was repeated until 17 envelopes of 4
bracelets were generated for subsequent issue to second users
(each a different individual). A further six bracelets handled
similarly were placed in a separate envelope as deposit controls.
Second users were provided with an information sheet, an
envelope with four worn bracelets, an activity data recording sheet
and four new small envelopes labelled 1–4 into which the worn
bracelets were to be placed upon completion of use. The 2nd
wearers were asked to start wearing all four bracelets at the same
time on the one wrist, and to record when they temporarily took
them off (e.g. when showering, sleeping) and put them back on.
They were also asked to wear the bracelets for different total time
periods. As close as possible these were to be of incremental
doubling of time (e.g. 5, 10, 20 and 40 h; or 1, 2, 4 and 8 d) and to
record on the data sheet the date and time they stopped wearing
each of the four bracelets. Buccal samples were collected from all
1st and 2nd wearers, from which their profiles were generated.
2.1.3. Test C
Individuals were asked to donate a personal object that could be
used by a second person for some time. When supplying an object
the owner was asked to confirm that they were the sole user of the
object, how long they had owned/used it and what was their
frequency and duration of its use. The object’s material composi-
tion and surface type were recorded.
Forty-six different objects (each from a different individual)
were issued to 2nd users (each a different individual) and returned
after use (Supplementary Table 1). Activity data sheets issued to
2nd users were as per Test A. Unlike with the items in Tests A and B
the initial amounts of DNA on these items were unknown. At the
completion of the object’s use by the 2nd user it was placed in
suitable clean packaging. Buccal samples were collected from all
original 1st owners and 2nd users from which their profiles were
generated.
 R.A.H. van Oorschot et al. / Forensic Science International: Genetics 8 (2014) 219–225
Supplementary material related to this article can be found, in
the online version, at doi:10.1016/j.fsigen.2013.10.005.
2.2. DNA collection, extraction, quantitation, amplification and
genotyping
2.2.1. Test A
The lid was removed from the pen and treated separately.
Wet + dry double swabbing with cotton swabs (150C, Copan, Italy),
was used to recover DNA for extraction. The whole external area of
each pen, excluding the writing tip and end tip, was targeted. The
external surface of each lid, excluding the end part of the clip, was
double swabbed separately. The two swabs of each item were
teased from the stick and placed in a spin basket within a 2 ml
centrifuge tube for DNA extraction.
2.2.2. Test B
Each bracelet was cut into small pieces of 3–6 mm using
cleaned scissors and forceps and these were placed into a 2 ml
centrifuge tube ready for DNA extraction.
2.2.3. Test C
Wet + dry double swabbing was used to collect DNA from hard
surfaces and tape lifting was used to collect DNA from soft, porous
objects.
2.2.4. Further processing
DNA was extracted from either the collection devices or directly
from the object using DNA IQ (Promega, USA) as per manufac-
turer’s instructions and routine procedures for similar casework
exhibits providing DNA extracts of 50 ml. The quantity of DNA in
each extract was determined using Quantifiler (Life Technologies,
USA) as per manufacturer’s instructions. Indicative Quantifiler
results below 23 pg/ml were utilised. Extractions and quantitations
were performed robotically on Biomek Nxp liquid handling
platforms (Beckman Coulter) The DNA from all samples, including
those with a negative quantitation result, was amplified to produce
profiles using AmpFlSTR Profiler Plus (Life Technologies, USA) in
25 ml reactions, run on an ABI PRISM 3130x genetic analyser and
analysed using GeneMapper ID v3.2 (Life Technologies, USA).
2.3. Data analyses
The genotypes of all 1st and 2nd users were known. The relative
profile percentage contributions of the 1st user/wearer, 2nd user/
wearer and unknown source were calculated for each test sample.
These values were calculated using relative RFU peak height
contributions at each locus. The peak height threshold was 50 RFUs.
Where the 1st and 2nd users were of the same sex Amelogenin was
excluded from the calculation, so too were any other loci sharing the
same genotype. Where an allele was shared at a locus the RFU
portions of that allele, derived from known contributors, were
determined using the RFU of the other alleles at that locus. The
relative profile contribution is the average of all informative loci.
2.3.1. Test A
To determine the effect of increased writing time on the DNA
yield and profile percentages contribution the pen data were
ranked from lowest to highest total writing time, and then grouped
into 5 writing time categories, and the average DNA yield and
profile percentage contribution by the 1st user, 2nd user and any
unknown source calculated for each category. It was assumed that,
as we requested, the lid was removed from the pen each time
writing occurred. To determine the effect of increased lid removal
over time on DNA yield and the profile percentages, lid data were
ranked based on the number of times the lid was removed from the
221
pen. Pens that were exposed to rubbing between hands by the 2nd
user and those that had not been used to write with by the 2nd user
were analysed separately from those that had been used just to
write with and left on a surface between writing activities. To
determine the effect, if any, of increasing DNA yield from the pens
on the profile percentage contribution from the 1st user, all pens
were ranked based on the DNA yield obtained and the average
profile percent contribution from the 1st user was calculated. This
procedure was repeated for the lids.
2.3.2. Test B
Of the 63 bracelets worn and returned for analysis, three, from
different groups, did not produce DNA profiles and were excluded
from further analyses.
A large number of alleles extraneous to the depositor were
noted on the deposit control bracelets. These alleles were referred
to as ‘1st wearer derived unknowns’. Alleles foreign to the 1st
wearer, 2nd wearer and ‘1st wearer derived unknowns’ were
referred to as ‘2nd wearer derived unknowns’.
To determine the effect of increased wear time on the DNA yield
and profile percentages, the bracelet data were ranked from lowest
to highest total wearing time. The data were grouped into 9 time
categories and the average DNA yields and profile percentage
contributions of the 1st wearer, 2nd wearer, 1st wearer derived
unknowns and 2nd wearer derived unknowns determined.
2.3.3. Test C
Objects sampled had been used by the owner for between 1
month and 10 years, and most (65%) were used every day or several
times a week. All items were owned and used solely by the owner.
120 samples were collected from 46 objects, with 108 returning
interpretable profiles.
3. Results
3.1. Test A: pens and lids
All five negative control pens and their corresponding lids gave
negative quantitation values and did not contain any allele when
profiled confirming that the pens and lids used in this study were
DNA-free before handling.
The five deposit control pens and their corresponding lids
yielded an average of 3.394 ng (range: 4.20–9.955 ng; S.D.: 0.638)
and 0.394 ng (range: 0.267–0.540 ng; S.D.: 0.118) respectively.
Each pen and lid returned a full profile of the 1st user. One of the
pens had one additional unknown allele, and another had two
other unknown alleles detected at different loci.
Of the 44 pens issued to 2nd users 37 had used the pen without
any obvious departures from normal use. Six other pens had been
rubbed between hands by the 2nd user (each returning high yields
of DNA) and one not used to write with, so these 7 were excluded
from evaluation of the effect of writing time on DNA persistence.
The pens were grouped into 5 writing time categories: 1–5 min
(n = 8), 6–30 min (n = 9), 31–60 min (n = 7), 61–90 min (n = 7) and
>90 min (n = 6). There was no significant difference in the average
quantity of DNA retrieved from the pens across the 5 groups. The
average yield of each group (ranging from 1.06 to 2.92 ng) was less
than the average of the deposit controls.
Of the corresponding lids, some were noted to have possibly been
sucked (one of which returned 24.35 ng of DNA and a profile only of
the 2nd user) or not returned leaving 30 for the analysis of DNA
quantity dependent on the number of times lids were removed and
replaced on the pen. The lids were grouped into 6 groups based on
the number of times the lid was removed and replaced: 1 (n = 12),
2 (n = 7), 3 (n = 3), 4 (n = 3), 5 (n = 2), 6 (n = 3). There was
variation in the quantity of DNA retrieved from the lids across the
222 R.A.H. van Oorschot et al. / Forensic Science International: Genetics 8 (2014) 219–225
Fig. 1. Average profile percentage contributions of the 2nd user, 1st user and
unknowns retrieved from pens per total writing time.
groups but without any apparent trends. Some lids provided more
than the deposit control lids and others less.
Fig. 1 shows the average profile percentage contribution from
the 1st user, 2nd user, and unknown source in each writing time
category. There appears to be an even mixture of profile percentage
contributions from the 1st and 2nd user when the total writing
time by the 2nd user was less than 30 min. After 30 min writing
time, the 2nd user forms the major contribution to the profile, and
after 90 min, the 1st user contributes an average of only 15% to the
mixture. There was some negative correlation between increased
writing time by the 2nd user and the profile percentage
contribution of the 1st user, but it was of borderline significance
(p = 0.052) in this relatively small sample set. However, when
comparing the shortest time writing group (1–5 min) to the
longest time writing group (>90 min) the difference between the
average DNA contributions by the 1st user was significant
(p < 0.01).
A small number of alleles of unknown origin were noted in 72%
of the pens analysed ranging from 1% to 9% of the total profile peak
RFUs. It appears that as writing time increases, the unknown
alleles’ percentage contribution decreases; however, this is
insignificant.
Fig. 2 shows that as the number of times a lid is removed from a
pen increases, the 2nd user’s profile percent contribution
Fig. 2. Average profile percentage contributions of the 2nd user, 1st user and
unknowns retrieved from pen-lids per number of lid removals.
increases, and the 1st user’s profile decreases. After one lid
removal the 2nd user forms the major profile percentage
contribution (average 58%). At 4 and 5 lid removals the 1st user
forms a very small percentage contribution (4% and 2% respec-
tively) and at 6 removals, the 1st user’s profile is not detectable,
and completely replaced by that of the 2nd user. Differences of
average profile contribution from the 1st user as the number of lid
removals increase is significant (p = 0.02).
Alleles of unknown origin were found in 53% of swabs from lids
analysed, ranging from 1% to 15% of the total profile peak RFUs. No
trend was observed in the percentage profile contribution of the
unknown source with the number of times the lid was removed.
Of the 6 pens reported to have been rubbed between the hands
by a 2nd user all pens and lids gave DNA quantities higher than the
average of the deposit controls. The profile of the 1st user was
present at a low percentage from three pens, and their
corresponding lids, but not in the other 3 where only the profile
of the 2nd user was obtained (data not shown).
3.2. Test B: bracelets
Of the 6 negative control bracelets three provided no DNA and
three very low quantities of DNA which provided either 1 or 2
alleles. The 6 deposit control bracelets yielded an average of
9.44 ng (range: 5.05–12.9 ng, S.D.: 3.42). All deposit control
bracelets provided full profiles of the 1st wearer. However, alleles
foreign to the 1st wearer were also found in each of the 6 bracelets.
The average profile percentage contributed by the 1st wearer was
85% (range: 81–88%, S.D.: 2.58) with the remainder being of foreign
origin. The number of foreign alleles per bracelet averaged 10.3
(range: 8–15). These extraneous alleles were similar across all 6
and none of those detected on the negative control bracelets were
found in the deposit controls.
The yields of DNA recovered from the 60 bracelets worn by a
2nd wearer ranged from 3.93 ng (worn for 10 min) to 217.5 ng
(worn for 16 days). The average amounts of DNA recovered from
each bracelet per time period worn by the 2nd wearer are shown in
Supplementary Fig. 1. 55 bracelets (92%) returned a higher DNA
quantity than the deposit control average of 9.44 ng. The 5
bracelets that returned a lower value than this were worn for a
short period (10–160 min). There is some positive relationship
between average DNA amounts and the lengths of time the
bracelet was worn by the 2nd wearer and a highly significant
difference between the DNA yields in each wearing time category
(p < 0.001).
Supplementary material related to this article can be found, in
the online version, at doi:10.1016/j.fsigen.2013.10.005.
Surprisingly, full profiles of the 1st wearer were recovered from
all but three bracelets. Of these latter three bracelets one was worn
for 16 days by the 2nd wearer and displayed three alleles derived
from the 1st wearer (at relatively low RFU) representing 2% of the
profile contribution. The other two bracelets, both worn for 8 days,
alleles from the 1st wearer were detected at 4 and 5 loci
contributing 7% and 12% of the total profile respectively.
Fig. 3 shows the percentages of DNA profiles derived from the
1st and 2nd wearer of the 60 bracelets worn by a 2nd wearer. As
the total time of bracelet wear by the 2nd wearer increases, the
percentage of the profile derived from the 1st wearer declines
significantly (p < 0.001). At 0–5 h of wear by the 2nd wearer, the
1st wearer still forms the major contribution to the profile. At 6–
28 h of wear by the 2nd wearer, there is a relatively equal
contribution. After 29 h of wear, the 2nd wearer provides the
majority contribution to the profile.
The number and overall contribution to the profile by unknown
alleles assumed to have been added by the 2nd wearer were low
with no clear trends of origin. Fig. 3 displays the average profile
 R.A.H. van Oorschot et al. / Forensic Science International: Genetics 8 (2014) 219–225
Fig. 3. Average profile percentage contributions of the 2nd wearer, 1st wearer and
unknowns derived from the 2nd and 1st wearer, retrieved from bracelets per
duration of wear by the 2nd wearer.
percentage contribution from unknown alleles identified in the
deposit controls (1st wearer) and those assumed to be derived
from the 2nd wearer. Similar to the trend seen in the known
profiles, the unknown alleles derived from the 1st wearer decrease
significantly as wearing time by the 2nd user increases (p < 0.001).
3.3. Test C: personal items
3.3.1. General
Details of the 120 areas of personal objects handled by a 2nd
user sampled and analysed, prior use by owner (1st user), use by
2nd user, DNA yields and profiles generated can be found in
Supplementary Table 1. 108 samples returned interpretable
profiles while the remaining 12 samples provided negative or
very low quantitation values and no profile for analysis and were
excluded from further analyses. The DNA collected from those
samples yielding DNA ranged from 0.01 ng to 91.5 ng. 28 samples
collected from porous substrates using the tape lift method
returned an average DNA yield of 31.37 ng. 92 samples collected
from non porous substrates via the double swab method returned
an average DNA yield of 2.57 ng (including 7 samples with 0 ng).
The majority (75.8%) of profiles generated from these items were
mixture profiles. Many (68.3%) included some alleles of unknown
origin.
There is a lot variation in the proportions that 1st owners and
2nd users contribute to the profiles in the 108 samples examined
and the duration of use by the 2nd user. There appears to be a slight
decline in contributions from the 1st owner and a slight increase in
contribution from the 2nd user as the duration of use by the 2nd
user increases, but these trends were statistically insignificant
(data not shown). Full profiles of the 1st user were recovered from
39.1% of non porous substrate samples and 78.6% of porous
substrate samples.
Alleles foreign to the 1st owner and 2nd user were found in
68.3% of the profiles. The highest portion of alleles of an unknown
origin observed within the profiles generated was 14%, the average
was 3.16, with 95% of samples displaying a portion of unknown
origin in the profile of 10%. A higher percentage of unknown
alleles were detected from samples collected from a porous
(average 4.6% profile contribution) compared to a non porous
substrate (average 2.7% profile contribution). These differences in
the number of alleles were significant (p < 0.05).
Several of the personal items and groups of similar items reveal
interesting results (Supplementary Table 1). Below we highlight
223
those from seven lanyards and associated ID holders and
summarise results of various other items.
3.3.2. Lanyards and ID holders
Seven lanyards (fabric) and associated ID holders (plastic) had
been worn constantly (9h/d for 5 d/wk) by the 1st owners for
various durations then worn similarly for different durations by
2nd wearers (Supplementary Figs. 2 and 3). The average quantity of
DNA recovered from the lanyards was 47.6 ng (S.D. 13.5) and full
profiles of the 2nd wearer were recovered from all samples. Full
profiles of the 1st owner were recovered from all samples except
for two used for 1 month by the 2nd wearer where a partial profile
of 12 alleles of the 1st owner was recovered. The profile
contribution derived from the 1st owner decreases as lanyard wear
by the 2nd wearer increases (Supplementary Fig. 2). Alleles of
unknown source were detected in all lanyards and represented
between 1% and 8% of the total profile. Similar results were
observed from the ID holders (Supplementary Fig. 3).
Supplementary material related to this article can be found, in
the online version, at doi:10.1016/j.fsigen.2013.10.005.
3.3.3. Summary results of other personal items
- Different sections of two well worn watches, with different types
of bands, exchanged for 10 d provided mainly full profiles from
the 1st owner and useful partial profiles from the 2nd user. Of the
four sections tested from each watch (outer and inner face and
band) the highest 1st wearer profile percentage contribution was
recovered from the outside of the bands (IDs 15–22, Supplemen-
tary Table 1).
- Whilst the internal surfaces and zippers of two well used purses,
exchanged for one month, provided full profiles of both the 1st
owner and 2nd user, only the profile of the 2nd user was
retrieved from their clips (IDs 27–34, Supplementary Table 1).
- Internal soles and shoe laces of two pairs of well worn sneakers
exchanged for 2 and 5 h respectively, provided full profiles of
both the 1st and 2nd user (IDs 113–120, Supplementary Table 1).
- For items such as; lighter, eye shadow case, USB stick, hair clip, lip
stick holder, lip gloss container and lid, pen and pen lid,
sunglasses nose and ear areas, perfume bottle lid the profile of
the 2nd user completely replaced that of the 1st owner after
various durations (including some for relatively short periods)
(Various IDs 35–78, Supplementary Table 1).
- The flap and inner rim of a cap well worn by a 1st owner and then
worn by a 2nd user for 12 h over 2 weeks provided full profiles of
both individuals with the 2nd user being the major contributor
(IDs 79–80, Supplementary Table 1).
See further details regarding these and other items in
Supplementary Table 1.
4. Discussion
All 6 pens that had been rubbed between hands by the 2nd user
and excluded from the comparison of pens based on their duration
of writing use, gave DNA yields higher than the deposit control
pens, with the 2nd user’s profile completely replacing that of the
1st user in 3 of these pens. This replacement of the 1st users’ DNA is
consistent with finding of Goray et al. [9] relating to the transfer of
DNA when touching hard non-porous substrates in a manner
where friction is involved.
It is noteworthy that most of the pens handled normally by a
2nd user provided less DNA than the deposit control pens held only
by the 1st user. This is possibly because the pens were transferred
to and from non-skin surfaces more times, consequently losing
some of their DNA to these other surfaces [9,10,12].
224 R.A.H. van Oorschot et al. / Forensic Science International: Genetics 8 (2014) 219–225
The opposite was observed with the bracelets in Test B where
the majority of bracelets worn by a 2nd wearer provided more DNA
than the deposit control bracelets. This difference between the two
tests is likely due to the longer contact the 2nd wearers had with
the bracelets compared to the 2nd users of the pens, as well as the
difference in substrate between the two items. This concurs with
what could be expected based on the findings of Wickenheiser [1]
and Goray et al. [9,13] that hard non-porous plastic has less
capacity to accumulate and retain deposited/transferred DNA than
soft rough surfaced porous fabric.
Substrate difference is also one of the reasons why the bracelets
returned more DNA than the pens even though the one dimensional
surface areas available for deposit of DNA were similar. A further
reason could be the difference in the method of DNA collection.
Swabs were used to collect DNA from the pens and the DNA then
extracted from the swabs whereas the bracelets were cut up and
DNA extracted directly from them. It has been demonstrated that
double swabbing does not collect all DNA present on a substrate
[14].
The presence of the 1st user’s DNA on pens, even on some held
for over 90 min by a 2nd user, may in part be because the 1st user
deposited their DNA over the whole of the pen while the 2nd users
handled the pen in a way that some areas remained untouched.
The variation observed within the various categories of samples
tested could be due to differences in shedder status of the 2nd
users [1,15,16] and the differences in the manner of contact [9].
The pressure and friction, when either removing or replacing the
pen lids, probably contributed to the rapid proportional decrease of
the 1st user’s profile and increase of the 2nd user’s profile [9]
observed in these samples.
The weak trend of reduced 1st user profile and increased 2nd
user profile over time observed in the collated results for all types
of personal items examined in Test C will in part relate to variation
in substrate and the size of area targeted for sampling. Of the
personal items examined, those with a porous substrate provided a
far greater proportion of full profiles of the original user than those
with a non porous substrate. This phenomenon concurs with the
findings of both Tests A and B.
As has been reported previously [1,8,17] the present study has
confirmed that depending on the type of object, the area in
question and the manner of handling the object, the DNA of the 2nd
user can be rapidly transferred to an object and replace the DNA of
a previous user. This is likely due to the simultaneous transfer from
the object to the handler reducing the total quantity of DNA
initially present, as well as the initial DNA becoming a smaller
portion of the total quantity of DNA present on the object after
being handled by a second person. One example of this was a USB
stick (hard flat plastic) used regularly by a single person for 5 yrs
and then used for just 1 min by a 2nd user. The 2nd user’s profile
was the only profile retrieved from it. This finding, however, is in
contrast to observations from items with rough surfaces, such as
the rubber thimble, which retained DNA of the original user, who
rarely used it, after a 2nd person used it actively for 1 min.
One can speculate that the reason for finding the highest 1st
wearer profile percentage contribution on the outer side of bands
of the two watches examined, rather than the inner side of the
bands and inner side of faces, is because this area has had the least
contact with the 2nd wearer. The higher profile percentage
contribution of the 1st wearer recovered from the inner band of the
metal link banded watch, relative to the straight leather banded
watch, is possibly because the former had ridges allowing DNA
containing material to build up over time.
Similar to the findings of the watches, the outer surface of the
straps of rubber sandals provided a higher percentage profile of the
1st wearer than the inner soles of sandals worn by a 2nd person.
Alleles of an unknown source were part of the profiles from the
inner soles of the sandals tested (Supplementary Table 1). The
presence of unknown alleles from swabs taken of feet has been
noted by others [18].
Alleles of an unknown source were found on the majority of
items examined in this study (i.e. pens, lids, bracelets and personal
items) and, when present, rarely exceeded more than 10% of the
total RFU of the profiles observed from a specific item. In
comparison Daly et al. [6] observed profiles in addition to the
depositors’ in a lower percentage of touched samples (37%, 25%
and 7.5% of single hand deposits on wood, fabric and glass
respectively). These samples however were of pre-cleaned objects
that had been touched only briefly once and included a high
portion of samples that yielded insufficient DNA to generate a
profile. The origin of the alleles of unknown source is likely to be
from individuals, and/or objects previously touched by others, that
the person or object being sampled has come into contact with.
As one would expect as the proportion of the total profile derived
from the 1st wearer of a bracelet in Test B declines as the wear by the
2nd wearer increases, so too did the proportion of unknown alleles
derived from the 1st wearer. However, the proportion of unknown
alleles derived from the 2nd wearers did not increase but remained
relatively constant. This finding is in part due to the variation in the
relative amounts of foreign DNA that an individual has on their
hands at any one time, and that in this experiment we observed an
average of several 2nd wearers, which happens to be less than the
relatively high level deposited by the one individual who deposited
their DNA on all the bracelets as the 1st wearer.
The results of this study provide insights into the likelihood of
obtaining the profile of a prior user of an object. They will assist
investigators decide which of the available relevant crime
associated objects, and which part(s) thereof, may best yield the
sought after information. This may improve the relative success
rate of collected samples, provide more probative information to
crime investigators as well as assist with the management of
resources (time, labour, consumables).
5. Conclusions
The degree of persistence of DNA from a prior user of an object
depends on the type of object, the substrate it is made of, the area
of the object targeted for sampling and the duration and manner of
contact by a subsequent user. Alleles of unknown sources should
be expected to be observed as a very minor component of the
profile generated from handled objects.
Greater knowledge of persistence will inform investigators
regarding the likelihood of detecting a profile of a particular
individual based on the type of object and its history, and assist with
identifying the best area(s) of an object to target for DNA sampling.
Acknowledgements
We would like to thank the volunteers who participated in this
study and Dr Kaye Ballantyne for assistance with the preparation of
the figures.
References
[1] R.A. Wickenheiser, Trace DNA: a review, discussion of theory, and application of
the transfer of trace quantities of DNA through skin contact, J. Forensic Sci. 47
(2002) 442–450.
[2] V. Castella, P. Mangin, DNA profiling success and relevance of 1739 contact stains
from casework, Forensic Sci. Int. Genet. Suppl. Ser. 1 (2008) 23–33.
[3] S.-A. Harbison, M. Fallow, D. Bushell, An analysis of the success rate of 908 trace
DNA samples submitted to the Crime Sample Database Unit in New Zealand, Aust.
J. Forensic Sci. 40 (2008) 49–53.
[4] J.J. Raymond, R.A.H. van Oorschot, P.R. Gunn, S.J. Walsh, C. Roux, Trace evidence
characteristics of DNA: a preliminary investigation of the persistence of DNA at
crime scenes, Forensic Sci. Int. Genet. 4 (2009) 26–33.
 R.A.H. van Oorschot et al. / Forensic Science International: Genetics 8 (2014) 219–225
[5] C. Dufva, A. Nilsson, Success rate of LT DNA analyses in casework, Forensic Sci. Int.
Genet. Suppl. Ser. 3 (2011) 271–272.
[6] D.J. Daly, C. Murphy, S.D. McDermott, The transfer of touch DNA from hands to
glass, fabric and wood, Forensic Sci. Int. Genet. 6 (2012) 41–46.
[7] J. Kenna, M. Smyth, L. McKenna, C. Dockery, S.D.S.D. McDermott, The recovery and
persistence of salivary DNA on human skin, J. Forensic Sci. 56 (2011) 170–175.
[8] J.J. Raymond, R.A.H. van Oorschot, S.J. Walsh, C. Roux, P.R. Gunn, Trace DNA and
street robbery: a criminalistic approach to DNA evidence, Forensic Sci. Int. Genet.
Suppl. Ser. 2 (2009) 544–546.
[9] M. Goray, R.J. Mitchell, R.A.H. van Oorschot, Investigation of secondary DNA
transfer of skin cells under controlled test conditions, Leg. Med. 12 (2010)
117–120.
[10] M. Goray, R.A.H. van Oorschot, R.J. Mitchell, Evaluation of multiple transfer of DNA
using mock case scenarios, Leg. Med. 14 (2012) 40–46.
[11] R.A.H. van Oorschot, M. Goray, E. Eken, R.J. Mitchell, Impact of relevant variables
on the transfer of biological substances, Forensic Sci. Int. Genet. Suppl. Ser. 2
(2009) 547–548.
225
[12] M. Goray, R.A.H. van Oorschot, R.J. Mitchell, DNA transfer within packaging:
potential for DNA loss and redistribution, Forensic Sci. Int. Genet. 6 (2012) 158–
166.
[13] M. Goray, E. Eken, R.J. Mitchell, R.A.H. van Oorschot, Secondary DNA transfer of
biological substances under varying test conditions, Forensic Sci. Int. Genet. 4
(2010) 62–67.
[14] B.C.M. Pang, B.K.K. Cheung, Double swab technique for collecting touched evi-
dence, Leg. Med. 9 (2007) 181–184.
[15] A. Lowe, C. Murray, J. Whitaker, G. Tully, P.P. Gill, The propensity of individuals to
deposit DNA and secondary transfer of low level DNA from individuals to inert
surfaces, Forensic Sci. Int. 129 (2002) 25–34.
[16] M. Phipps, S.F. Petricevic, The tendency of individuals to transfer DNA to handled
items, Forensic Sci. Int. 168 (2007) 162–168.
[17] R.A.H. van Oorschot, M.K. Jones, DNA fingerprints from fingerprints, Nature 387
(1997) 767.
[18] J. Bright, S.F. Petricevic, Recovery of trace DNA and its application to DNA profiling
of shoe insoles, Forensic Sci. Int. 145 (2004) 7–12.