Highlights
- •Partitioning tapelifts into multiple sections for numerous direct PCRs was tested.
- •Substrate type and donor were significant for DNA profiling success and LR support.
- •Intra- tapelift samples clustered more closely than inter- tapelift samples.
- •Profile to profile comparisons showed high similarity for intra- tapelift samples.
- •Direct PCR and extraction portions taken from a single tapelift showed concordance.
Abstract
Although a version of direct PCR is implemented in forensic laboratories for reference
material, its incorporation into workflow for the analysis of touch DNA, as a form
of latent DNA, from casework exhibits is not. In addition to concerns about increased
sensitivity causing more complex mixtures or the generation of more genetic data implicating
an individual superfluous to the context of the alleged event, the complete use of
the collected sample in the PCR as template has meant that there is no possibility
for data reproducibility when needed. Here it is proposed that the use of tapelifts
in touch DNA collection can facilitate replicate direct PCR analysis from a single
sample allowing the sample to be re-tested. If all portions of the tapelift result
in profiles with allelic and likelihood ratio concordance, these sub-samples may be
accepted as technical replicates, thus meeting any accreditation guideline requirements.
Furthermore, we assess the use of a single tapelift for both direct PCR and extraction-based
PCR workflows to illustrate the potential for benefits of both systems to be facilitated.
DNA was deposited by three donors onto six substrates with five sample replicates
of each condition. Separation of each tapelift into three portions for three direct
PCRs ensued using VeriFiler™ Plus. Separation of single tapelifts into three direct
PCRs showed no statistical difference in donor allele calls or RFU, or subsequent
LRs associated with their profiles. Comparison of profiles within the single tapelift
showed more similarity, with high mixture-to-mixture match likelihoods, than when
these sub-samples were compared with profiles generated from other samples. This allows
each sub-sample taken from the tapelift to be considered as technical replicates.
For dual workflow facilitation assessment, one donor deposited DNA through touch onto
six substrates with five research replicates of each. Separation of single tapelifts
into two portions, one for direct PCR and the retention and use of the remaining portion
for extraction and subsequent PCR, showed no significant difference in allelic yield
and subsequent donor comparison LRs. Comparison of deconvoluted profiles produced
from a single tapelift showed high mixture-to-mixture match likelihoods, supporting
DNA donor concordance. This indicates that removing a portion of a tapelift for direct
PCR amplification, while processing the remainder through standard processes, allows
increased sensitivity through direct PCR while offering the preparation of an eluate
suitable for repeated analyses.
Keywords
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Article info
Publication history
Published online: December 10, 2021
Accepted:
December 8,
2021
Received in revised form:
October 28,
2021
Received:
September 7,
2021
Identification
Copyright
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