Understanding the factors that may impact the transfer, persistence, prevalence and recovery of DNA (DNA-TPPR), and the availability of data to assign probabilities to DNA quantities and profile types being obtained given particular scenarios and circumstances, is paramount when performing, and giving guidance on, evaluations of DNA findings given activity level propositions (activity level evaluations). In late 2018 and early 2019, three major reviews were published on aspects of DNA-TPPR, with each advocating the need for further research and other actions to support the conduct of DNA-related activity level evaluations. Here, we look at how challenges are being met, primarily by providing a synopsis of DNA-TPPR-related articles published since the conduct of these reviews and briefly exploring some of the actions taken by industry stakeholders towards addressing identified gaps. Much has been carried out in recent years, and efforts continue, to meet the challenges to continually improve the capacity of forensic experts to provide the guidance sought by the judiciary with respect to the transfer of DNA.
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Considering activity level propositions in the evaluation of forensic biology findings is becoming more common place. There are increasing numbers of publications demonstrating different transfer mechanisms that can occur under a variety of circumstances. Some of these publications have shown the possibility of DNA transfer from site to site on an exhibit, for instance as a result of packaging and transport. If such a possibility exists, and the case circumstances are such that the area on an exhibit where DNA is present or absent is an observation that is an important diagnostic characteristic given the propositions, then site to site transfer should be taken into account during the evaluation of observations. In this work we demonstrate the ways in which site to site transfer can be built into Bayesian networks when carrying out activity level evaluations of forensic biology findings. We explore the effects of considering qualitative vs quantitative categorisation of DNA results. We also show the importance of taking into account multiple individual’s DNA being transferred (such as unknown or wearer DNA), even if the main focus of the evaluation is the activity of one individual.
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Adhesive tape is a common piece of evidence that can contain a myriad of traces. Due to its adhesive properties, adhesive tape can potentially collect traces unrelated to the crime or relocate crime-relevant traces. This secondary transfer of traces can have crucial implications for the evaluation at the activity level. Therefore, this study investigated the secondary transfer of DNA between layers of adhesive tape and tape and other case- and laboratory-relevant substrates. A drop of diluted blood was deposited on different primary substrates (i.e. duct tape, metal, plastic, textile, nitrile gloves). Subsequently, the primary substrate was brought into contact with a secondary substrate, and DNA was collected from both surfaces to measure transfer rates. The highest transfer rates were detected between the adhesive side of the tape and plastic, whereas the lowest transfer rates were detected between the adhesive side and textile. It was shown that the adhesive readily collects DNA from plastic and nitrile gloves commonly used in the laboratory, which highlights the importance of working with DNA-free materials. Therefore, this study demonstrated the need for caution when interpreting traces on adhesive tapes, always taking possible situations of secondary transfer into account.
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A large, recently published, inter-laboratory study by the ReAct group has shown that there is considerable variability in DNA recovery that exists between forensic laboratories. The presence of this inter-laboratory variability presents issues when one laboratory wishes to carry out an evaluation and needs to use the data produced by another laboratory. One option proposed by the ReAct group is for laboratories to carry out a calibration exercise so that appropriate adjustments between laboratories can be made. This will address some issues, but leave others unanswered, such as how to make use of the decades of transfer and persistence data that has already been published. In this work we present a method to utilise data produced in other laboratories (whether it provides DNA amounts or a probability of transfer) that takes into account inter-laboratory variability within an evaluation. This will allow evaluations to continue, without calibration data, and ensures that the strength of findings is appropriately represented. In this paper we discuss complicating factors with the various ways in which previous data has been reported, and their limitations in supporting probability assignments when carrying out an evaluation. We show that a combination of producing calibration information for new data (as suggested by the ReAct group) and development of strategies where calibration data is not available will provide the best way forward in the field of evaluations given activities.
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In this study, we assessed to what extent data on the subject of TPPR (transfer, persistence, prevalence, recovery) that are obtained through an older STR typing kit can be used in an activity-level evaluation for a case profiled with a more modern STR kit. Newer kits generally hold more loci and may show higher sensitivity especially when reduced reaction volumes are used, and this could increase the evidential value at the source level. On the other hand, the increased genotyping information may invoke a higher number of contributors in the weight of evidence calculations, which could affect the evidential values as well. An activity scenario well explored in earlier studies [1,2] was redone using volunteers with known DNA profiles. DNA extracts were analyzed with three different approaches, namely using the optimal DNA input for (1) an older and (2) a newer STR typing system, and (3) using a standard, volume-based input combined with replicate PCR analysis with only the newer STR kit. The genotyping results were analyzed for various aspects such as percentage detected alleles and relative peak height contribution for background and the contributors known to be involved in the activity. Next, source-level LRs were calculated and the same trends were observed with regard to inclusionary and exclusionary LRs for persons who had or had not been in direct contact with the sampled areas. We subsequently assessed the impact on the outcome of the activity-level evaluation in an exemplary case by applying the assigned probabilities to a Bayesian network. We infer that data from different STR kits can be combined in the activity-level evaluations.
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To accelerate differentiation between Staphylococcus aureus and Coagulase Negative Staphylococci (CNS), this study aimed to compare six different DNA extraction methods from 2 commonly used blood culture materials, i.e. BACTEC and Bact/ALERT. Furthermore, we analyzed the effect of reduced blood culture times for detection of Staphylococci directly from blood culture material. A real-time PCR duplex assay was used to compare 6 different DNA isolation protocols on two different blood culture systems. Negative blood culture material was spiked with MRSA. Bacterial DNA was isolated with: automated extractor EasyMAG (3 protocols), automated extractor MagNA Pure LC (LC Microbiology Kit MGrade), a manual kit MolYsis Plus, and a combination between MolYsis Plus and the EasyMAG. The most optimal isolation method was used to evaluate reduced bacterial culture times. Bacterial DNA isolation with the MolYsis Plus kit in combination with the specific B protocol on the EasyMAG resulted in the most sensitive detection of S.aureus, with a detection limit of 10 CFU/ml, in Bact/ALERT material, whereas using BACTEC resulted in a detection limit of 100 CFU/ml. An initial S.aureus load of 1 CFU/ml blood can be detected after 5 hours of culture in Bact/ALERT3D by combining the sensitive isolation method and the tuf LightCycler assay.
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In sexual assault cases, the retrieved DNA quantity and sampling location from the victim’s underwear may provide valuable information for activity level evaluative reporting. DNA can transfer from site to site on an exhibit, or be lost within packaging, complicating interpretation. Experiments are needed to investigate these factors. This preliminary study compared two cleaning methods to prepare undergarments for such experimentation: hand-washing with warm water and washing with bleach before rinsing. Results show a significantly lower quantity of DNA on washed underwear using both methods. Warm-water hand-washing, the more straightforward method, was selected for further experimentation.
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There appears to be some hesitation within the forensic biology community to formally evaluate and report on findings given activity level propositions. This hesitance in part stems from concerns about the lack of relevant data on the dynamics of biological traces and doubt about the relevance of such expert opinions to the trier of fact. At the Netherlands Forensic Institute formal evaluative opinions on the probability of case findings given propositions at the activity level are provided since 2013, if requested by a mandating authority. In this study we share the results from a retrospective analysis of 74 of such requests. We explore which party initiates requests, the types of cases that are submitted, the sources of data being used to assign probabilities to DNA transfer, persistence, prevalence and recovery (TPPR) events, the conclusions that were drawn by the scientists, and how the conclusions were used by the courts. This retrospective analysis of cases demonstrates that published sources of data are generally available and can be used to address DNA TPPR events in most cases, although significant gaps still remain. The study furthermore shows that reporting on forensic biology findings given activity level propositions has been generally accepted by the district and appeal courts, as well as the other parties in the criminal justice system in the Netherlands.
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Plasmid-mediated dissemination of antibiotic resistance among fecal Enterobacteriaceae in natural ecosystems may contribute to the persistence of antibiotic resistance genes in anthropogenically impacted environments. Plasmid transfer frequencies measured under laboratory conditions might lead to overestimation of plasmid transfer potential in natural ecosystems. This study assessed differences in the conjugative transfer of an IncP-1 (pKJK5) plasmid to three natural Escherichia coli strains carrying extended-spectrum beta-lactamases, by filter mating. Matings were performed under optimal laboratory conditions (rich LB medium and 37°C) and environmentally relevant temperatures (25, 15 and 9°C) or nutrient regimes mimicking environmental conditions and limitations (synthetic wastewater and soil extract). Under optimal nutrient conditions and temperature, two recipients yielded high transfer frequencies (5 × 10–1) while the conjugation frequency of the third strain was 1000-fold lower. Decreasing mating temperatures to psychrophilic ranges led to lower transfer frequencies, albeit all three strains conjugated under all the tested temperatures. Low nutritive media caused significant decreases in transconjugants (−3 logs for synthetic wastewater; −6 logs for soil extract), where only one of the strains was able to produce detectable transconjugants. Collectively, this study highlights that despite less-than-optimal conditions, fecal organisms may transfer plasmids in the environment, but the transfer of pKJK5 between microorganisms is limited mainly by low nutrient conditions.
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In this case study, we want to gain insights into how residents of three municipalities communicate about the new murder scenario of the cold case of Marianne Vaatstra and the possibility of a large-scale DNA familial searching. We investigate how stakeholders shape their arguments in conversation with each other and with the police. We investigate the repertoires that participants use to achieve certain effects in their interactions with others in three focus groups. The results show that the analyzed repertoires are strong normative orientated. We see two aspects emerge that affect the support for large-scale DNA familial searching. These are: 1. Cautious formulations: respondents showed restraint in making personal judgments and often formulated these on behalf of others. Participants would not fully express themselves, but adjusted to what seemed the socially desirable course. 2. Collective identity: respondents focused on the similarities between themselves and the needs, interests, and goals of other participants. Participants also tried in a discursive way to convince each other to participate in the large-scale familial searching. These two major discursive activities offered the communication discipline guidance for interventions into the subsequent communication strategy.
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