Touch Deposits Research Articles

The impact of substrate characteristics on the collection and persistence of biological materials, and their implications for forensic casework

This study assessed the level of nucleic acid persistence on the substrate pre-, and post-swabbing, in order to assess whether biological materials (touch, saliva, semen, and blood) are collected differently depending on the substrate characteristics. A total of 48 samples per deposit and substrate variety (n = 384) were assessed by tracking the persistence of nucleic acid using Diamond™ Nucleic Acid Dye (DD) staining and Polilight photography. The number of DD nucleic acid fluorescent complexes formed post-staining were counted (fluorescent count) and in conjunction with the fluorescence signal intensity (DD nucleic acid complex accumulation) used to estimate the level of nucleic acid persistence on substrates. Touch deposits have shown to be the most persistent deposit with strong adhesion capabilities on both substrate verities. Saliva displayed a higher persistence than semen and/or blood. Semen displayed a high collection efficiency as well as a high fluorescence signal intensity. Blood displayed a low persistence on both substrates with a superior collection efficiency that may also indicate a higher probability to become dislodged from surfaces given a particular activity. Our research has shown that the persistence and recovery of biological deposits is not only measurable but more importantly, may have the potential to be estimated, as such, may build an understanding that can provide valuable guidance for collection efficiency evaluations, and the assessing of the probability of particular profiles, given alternate propositions of means of transfer occurring.

Determining the number and size of background samples derived from an area adjacent to the target sample that provide the greatest support for a POI in a target sample

When sampling an item or surface for DNA originating from an action of interest, one is likely to collect DNA unrelated to the action of interest (background DNA). While adding to the complexity of a generated DNA profile, background DNA has been shown to aid in resolving the genotypes of contributors in a targeted sample, and where references of donors to the background DNA are not available, strengthen the LR supporting a person of interest contributing to the targeted sample. This is possible thanks to advances in probabilistic genotyping, where forensic labs are able to deconvolute complex DNA profiles to obtain lists of genotypes and their associated weights. Coupled with DBLR™, one can then compare multiple evidentiary profiles to each other to determine the contribution of common, but unknown, contributors. Here, we consider factors associated with taking background samples and whether one should collect multiple background samples that all relate to a single target sample, or if one should collect larger background samples rather than smaller samples. Background samples consisted of DNA accumulated on the items primarily by one or both occupants of a single household, while targeted samples were generated from touch deposits, or saliva deposits that had been left to air dry. Samples were collected from areas of various sizes, consisting of only the background, the target and the background directly beneath it, and the target and additional surrounding background. A broad range of DNA quantities were recovered, with larger background samples (400 cm2) yielding significantly more DNA than smaller background samples (30 cm2). Significant differences in DNA quantities between target samples were not observed. Generated DNA profiles were interpreted using STRmix™ and DBLR™, and where there was support for a common donor between the background and target sample, pairwise comparisons were performed to observe the effect on the LR supporting the target DNA donor contributing to the targeted sample when conditioning on one (or two) common donor between the targeted sample and 1–8 background samples. Multiple background samples gave significantly higher LRs compared to a single background sample, the larger sampled background area resulted in larger LR gains than the smaller areas, and four or more background samples reduced LR variability considerably. Here we provide recommendations for the minimum and ideal number of additional background samples that should be collected, and that several smaller samples may be more beneficial than a single larger sample.

How changes to the substrate’s physical characteristics can influence the deposition of touch and salivary deposits

An in-depth study into the physical substrate characteristics such as substrate surface roughness, topography, and physicochemical characteristics like wettability and surface free energy (SFE) was conducted to investigate the impact on the deposition and adherence of touch and salivary deposits on aluminium and polypropylene. A robust protocol was established to generate a set of substrates with a controlled linear surface roughness range (0.5–3.5 µm) in order to identify the impact of surface roughness on DNA transfer, persistence, prevalence, and recovery (DNA-TPPR). The polypropylene substrate was shown to produce fibres when artificially roughened, becoming more prominent at a higher surface roughness range, and has shown to have a direct impact on the distribution of salivary and touch deposits. At the low to moderate surface roughness range 0.5–2.0 µm, salivary and touch deposits have generally shown to follow the topographical features of the substrate they were deposited on, before a plateau of the surface roughness measure on the deposit was observed, indicating that a saturation point was reached and the grooves in the substrate were beginning to fill. Touch deposits have shown to maintain a consistent deposition height pre-surface roughness threshold, irrespective of substrate surface roughness while the deposition height of salivary deposits was heavily influenced by substrate surface roughness and topography. The substrate SFE, wettability, hydrophobicity, and the surface tension of the deposit was shown to drive the adhesion properties of the saliva and touch deposits on the respective substrates, and it was observed that this may be of importance for the improvement of the current DNA-TPPR understanding, DNA sampling protocols, and DNA transfer considerations within casework.

Reset Required: The Euro-area Crisis Management and Deposit Insurance Framework

Abstract The crisis management and deposit insurance (CMDI) framework in the euro area requires a reset. This article explains why and proposes a new framework. This could start as early as 1 January 2024 when significant institutions in the euro area will have met requirements to have enough subordinated obligations outstanding to recapitalize the bank if it were to fail. The proposed framework has four components: a single lender of last resort (the European Central Bank); a single presumptive path for resolution (exit via the use of bail-in to facilitate orderly liquidation of the failed bank by the Single Resolution Board under a solvent wind-down strategy); an investor of last resort in a bank’s gone-concern capital (its national deposit guarantee scheme); and a Single Deposit Guarantee Scheme (the Single Resolution Fund with a backstop from the European Stability Mechanism). Together, these measures would limit forbearance, assure bail-in did not touch deposits, promote competition, limit recourse to taxpayer money, standardize resolution procedures for all banks, complete Banking Union and guarantee that a euro of covered deposits would remain a euro, all without forcing national deposit guarantee schemes to reinsure one another. Last but not least the proposed framework would promote financial stability.

Technical Note: Lysis and purification methods for increased recovery of degraded DNA from touch deposit swabs

Touch deposits are a routine yet challenging sample type in forensic casework and research. Recent work investigating their contents has indicated corneocytes to be the major cellular constituent while cell-free DNA is present at significant levels. Prolonged incubation including a reducing agent such as DTT has been shown to lyse corneocytes; a plasma cfDNA recovery kit which targets shorter DNA fragments has been demonstrated to improve cfDNA recovery from hand rinses. Herein these methods are combined and tested on mock casework touch deposit swabs from communal surface areas. Both fluorescence- and qPCR-based quantification methods are used and their results compared to query DNA degradation levels. Both proposed lysis and purification methods demonstrate increased recovery of DNA detectable with fluorescence quantification and some additional alleles at short loci, indicating high levels of fragmented DNA in these samples

How many cells are required for successful DNA profiling?

Through advances in fluorescent nucleic acid dye staining and visualisation, targeted collection of cellular material deposited, for example by touch or within a saliva deposit, is possible. In regard to the potential evidentiary value of the deposit the questions remain: ‘How many cells are required to generate an informative DNA profile?’; ‘How many visualised corneocytes within a touch deposit compared to typical nucleated cells are required in order to achieve successful DNA profiling?’.Diamond TM Nucleic Acid Dye (DD) staining of cellular material, and subsequent visualisation utilising portable fluorescence microscopy, was performed for touch and saliva samples to target defined numbers of cells for collection, by swab and tapelift, and subsequent processing via direct PCR and PCR post-extraction. The resulting DNA quantification data and alleles generated within subsequent DNA profiles could be correlated to the number of cells initially collected to determine cellular threshold requirements for DNA profile generation for each workflow.Full profiles were consistently generated using direct PCR when the template was ≥40 buccal cells collected by either a swab or tapelift. By contrast ≥800 corneocytes collected by swabbing or ≥4,000 corneocytes collected by a tapelift were required to generate same number of STR alleles from touch samples. When samples were processed through a DNA extraction workflow, ≥80 buccal cells were required to generate full profiles from both swab and tapelift, while touch samples required ≥4,000 corneocytes collected by a swab and >8,000 corneocytes collected by a tapelift. The data presented within this study allow for informative sample triage and workflow decisions to be made to optimise STR amplification based on the presence and visual quantification of stained cellular material.

Corneocyte lysis and fragmented DNA considerations for the cellular component of forensic touch DNA

DNA deposited by individuals’ hands is a routine part of forensic analysis, yet little is understood about the precise cellular contents left by handling. “Dead” skin cells known as corneocytes make up the majority of the cellular material left in touch deposits by people’s hands but are known to lack nuclei, making their DNA content ambiguous. Here we measure DNA released from anucleate corneocytes following various lysis methods to determine how much DNA may be present in these cells and how best to recover it from inside the cornified envelope. We demonstrate that enhanced lysis methods using a reducing agent and longer incubation may be valuable for hand deposit samples. Corneocyte DNA can be characterized as highly degraded based on the quantification, STR profiling and fluorescence microscopy of the cells from freshly washed hands. Purification to target shorter DNA fragments is demonstrated. DNA from the washed corneocyte cells is shown to constitute the majority of recoverable DNA with these methods. We consider the use of new methods adapted to cornified cells and fragmented DNA for future research into this sample type

Exploration of cell-free DNA (cfDNA) recovery for touch deposits

Although touch deposit DNA is widely used in forensic casework, its cellular and acellular contents and their biological origins are poorly understood. There is evidence that the cell-free component of DNA deposited by handling may contribute substantial genetic information; however, most research into touch DNA recovery does not separate cellular and cell-free fractions or seek to characterize their contents. This work is an important early step in developing methods to isolate the cfDNA from biological material deposited by handling. Size-filtration as a separation technique was determined to be prone to DNA loss, even on optimized control samples of pure ladder DNA. Centrifugal separation was optimized to determine minimum speed and time required to reliably remove all cellular debris from the material collected by rinsing donor hands. To determine if the centrifugal force risked rupturing shed corneocyte cells and releasing cellular DNA into the supernatant, DNA levels were measured, and cells were visualized microscopically before and after centrifugation of hand rinses. Heated buccal cells were used as a positive control to demonstrate cell rupture would be detected with these methods. Following the determination of a suitable separation technique, an investigation into purification methods for cfDNA was conducted. DNA recovery using three kits for plasma cfDNA, one for PCR clean-up and one for genomic DNA were assessed on both ladder DNA to simulate cfDNA fragments and on collected hand deposit supernatants from both unwashed and washed hands. Purification methods designed for recovery of short DNA fragments from plasma yielded the highest recovery percentage across sample types, with BioChain cfPure performing the best. Donors’ hands were shown to shed high levels of cfDNA, which were better recovered with a method for short fragments than with a traditional genomic technique often used on touch DNA samples.

Illuminating touch deposits through cellular characterization of hand rinses and body fluids with nucleic acid fluorescence

Forensic DNA typing from touched or handled items in routine casework is increasing as the sensitivity of detection techniques improves. Our understanding of the cellular/acellular content of touch deposits and the origins of the DNA therein is still limited. This work explores the cellular content of rinses from washed and unwashed hands, as well as saliva, nasal and eye washes which could be sources of transferred DNA onto hands. Flow cytometry and microscopic examination were used to detect granularity, size and nucleic acid fluorescence data. Cellular content did not vary significantly within an individual, although some differences were observed between donors. Saliva contained populations of nucleated epithelia as well as smaller cells and debris, all positive for DNA. Hand rinses consisted almost entirely of anucleate corneocytes, many of which also stained positive for nucleic acids. These data raise questions about shed corneocyte DNA content previously assumed to be negligible.

Touch DNA: impact of handling time on touch deposit and evaluation of different recovery techniques: An experimental study

“Touch DNA” is DNA obtained from biological material transferred from a donor to an object or a person during physical contact. This particular kind of evidence could play an essential role in forensic laboratory work and is considered an important tool for investigators. Even though the principal aspects of “Touch DNA” have been extensively studied, to date, there are few reports in the research field of DNA retrieval from garments that have been worn. This study aimed to investigate the “handling time”, analyzing particularly the minimum contact time required to deposit a sufficient amount of DNA on a garment to produce an interpretable profile of the “handler”. Moreover, three different sampling techniques were compared (“dry swab”, “cutting out”, and “adhesive tape”) with the aim of defining the technique that guarantees the best recovery of the three methods tested. Analyzing the data of this experimental model, a “handling time” of two seconds is enough to release sufficient DNA on to a garment to obtain a complete profile. Moreover, this study demonstrated that when targeting for foreign DNA, the sample area should be narrowed down as much as possible to the smallest area possible to maximize target DNA recovery.

Evaluation of tapelifting as a collection method for touch DNA

The use of tapelifting for collection of touch DNA from fabrics is routine in many jurisdictions. However, there is a paucity of data relating to the effectiveness of different types of tapes for tapelifting, the amount of tapelifting required to generate a useful profile, and whether or not tapelifting is more effective than swabbing from various substrates. This research investigates these questions by comparing two tapes of different adhesive strength currently used in forensic casework (Scotch Magic tape and Scenesafe FAST minitapes), for sampling from touch deposits on four different fabrics-cotton flannelette, cotton drill woven fabric, polyester/cotton plain woven fabric and polyester strapping. Touch DNA was deposited on four replicates of each substrate. Separate areas of each substrate replicate were sampled, either by taping with one of the two tapes or by wet/dry swabbing with cotton swabs. Tape was applied over the defined sampling area once or repeatedly for various numbers of applications. DNA was extracted, quantified and profiled from all tape and swab samples as well as the corresponding sampled substrates. Significantly more DNA was extracted, and a higher proportion of alleles detected, from Scenesafe FAST tape than from Scotch Magic tape. The amount of DNA and number of donor alleles detected generally increased as the tape was reapplied to the surface, although a threshold of collection was seen for both types of tape. For two out of four substrates, taping with Scenesafe FAST collected more DNA than swabbing and, for three substrates, generated a greater median number of donor alleles. There was no significant difference in numbers of alleles between swabbing and taping from flannelette. Based on these findings, it is recommended that a tape with stronger adhesion (such as Scenesafe FAST tapelifters) is generally preferable; that more than one application of tape is suggested (however, increasing the amount of times the area is sampled can diminish collection efficiency); and that there is an advantage using tapelifting rather than swabbing for fabrics unless, such as with flannelette, there are many loose fibres easily removed during the sampling process.