Editorial on the recommendations of the DNA commission of the ISFG on the interpretation of mixtures.
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DNA↗
Biomedical subjects
Peter Gill
Publications and source records attributed to Peter Gill.
LoComatioN: a software tool for the analysis of low copy number DNA profiles.
Previously, the interpretation of low copy number (LCN) STR profiles has been carried out using the biological or 'consensus' method-essentially, alleles are not reported, unless duplicated in separate PCR analyses [P. Gill, J. Whitaker, C. Flaxman, N. Brown, J. Buckleton, An investigation of the rigor of interpretation rules for STRs derived from less than 100 pg of DNA, Forens. Sci. Int. 112 (2000) 17-40]. The method is now widely used throughout Europe. Although a probabilistic theory was simultaneously introduced, its time-consuming complexity meant that it could not be easily applied in practice. The 'consensus' method is not as efficient as the probabilistic approach, as the former wastes information in DNA profiles. However, the theory was subsequently extended to allow for DNA mixtures and population substructure in a programmed solution by Curran et al. [J.M. Curran, P. Gill, M.R. Bill, Interpretation of repeat measurement DNA evidence allowing for multiple contributors and population substructure, Forens. Sci. Int. 148 (2005) 47-53]. In this paper, we describe an expert interpretation system (LoComatioN) which removes this computational burden, and enables application of the full probabilistic method. This is the first expert system that can be used to rapidly evaluate numerous alternative explanations in a likelihood ratio approach, greatly facilitating court evaluation of the evidence. This would not be possible with manual calculation. Finally, the Gill et al. and Curran et al. papers both rely on the ability of the user to specify two quantities: the probability of allelic drop-out, and the probability of allelic contamination ("drop-in"). In this paper, we offer some guidelines on how these quantities may be specified.
Alleles↗
New multiplexes for Europe-amendments and clarification of strategic development.
Recently, the ENFSI/EDNAP groups issued advice on the design of the next generation of STR multiplexes in order to encourage standardisation within Europe. As the result of collaborative experimentation within the EDNAP group, we demonstrated that the low molecular weight STRs had substantial benefits to detect degraded samples. We subsequently recommended adoption of three new mini-STR loci to improve the success rate of degraded DNA markers, concurrent with the reduction in size of the existing STR markers in current use. This also improves the discriminating power of the system which is important to improve the power of national DNA databases. Subsequent discussions have occurred with manufacturers and members of the ENFSI/EDNAP groups. Because significant time and investment is required to develop new multiplexes of 13+ STR loci, manufacturers indicated that it would be preferable to adopt a staged approach. Two differing, but parallel strategies have now emerged. The first strategy employs a 13 STR loci multiplex incorporating three mini-STRs into the current multiplex test. The second strategy employs a multiplex of six high molecular weight STRs (in current use), modified to provide smaller amplicons combined with an additional two loci of high discriminating power. Eventually, the two strategies will converge to provide a single multiplex of 15 STR loci. The process will be guided by the ENFSI/EDNAP groups.
DNA Fingerprinting↗
The evolution of DNA databases--recommendations for new European STR loci.
Following a recent meeting by the ENFSI and EDNAP groups on the 4-5 April, 2005, in Glasgow, UK, it was unanimously agreed that the process of standardization within Europe should take account of recent work that unequivocally demonstrated that chance of obtaining a result from a degraded sample was increased when small amplicons (mini-STRs) were analysed. Consequently, it was recommended that existing multiplexes are re-engineered to enable small amplicon detection, and that three new mini-STR loci with alleles <130 bp (D10S1248, D14S1434 and D22S1045) are adopted as universal. This will increase the number of European standard Interpol loci from 7 to 10.
DNA Fingerprinting↗
DNA as evidence--the technology of identification.
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Criminal Law↗
PENDULUM--a guideline-based approach to the interpretation of STR mixtures.
Several years ago, a theory to interpret mixed DNA profiles was proposed that included a consideration of peak area using the method of least squares. This method of mixture interpretation has not been widely adopted because of the complexity of the associated calculations. Most reporting officers (RO) employ an experience and judgement based approach to the interpretation of mixed DNA profiles. Here we present an approach that has formalised the thinking behind this experience and judgement. This has been written into a computer program package called PENDULUM. The program uses a least squares method to estimate the pre-amplification mixture proportion for two potential contributors. It then calculates the heterozygous balance for all of the potential sets of genotypes. A list of "possible" genotypes is generated using a set of heuristic rules. External to the programme the candidate genotypes may then be used to formulate likelihood ratios (LR) that are based on alternative casework propositions. The system does not represent a black box approach; rather it has been integrated into the method currently used by the reporting officers at the Forensic Science Service (FSS). The time saved in automating routine calculations associated with mixtures analysis is significant. In addition, the computer program assists in unifying reporting processes, thereby improving the consistency of reporting.
Algorithms↗
A graphical simulation model of the entire DNA process associated with the analysis of short tandem repeat loci.
The use of expert systems to interpret short tandem repeat DNA profiles in forensic, medical and ancient DNA applications is becoming increasingly prevalent as high-throughput analytical systems generate large amounts of data that are time-consuming to process. With special reference to low copy number (LCN) applications, we use a graphical model to simulate stochastic variation associated with the entire DNA process starting with extraction of sample, followed by the processing associated with the preparation of a PCR reaction mixture and PCR itself. Each part of the process is modelled with input efficiency parameters. Then, the key output parameters that define the characteristics of a DNA profile are derived, namely heterozygote balance (Hb) and the probability of allelic drop-out p(D). The model can be used to estimate the unknown efficiency parameters, such as pi(extraction). 'What-if' scenarios can be used to improve and optimize the entire process, e.g. by increasing the aliquot forwarded to PCR, the improvement expected to a given DNA profile can be reliably predicted. We demonstrate that Hb and drop-out are mainly a function of stochastic effect of pre-PCR molecular selection. Whole genome amplification is unlikely to give any benefit over conventional PCR for LCN.
Computer Graphics↗
Ongoing controversy over Romanov remains.
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Bayes Theorem↗
STR analysis of artificially degraded DNA-results of a collaborative European exercise.
Degradation of human DNA extracted from forensic stains is, in most cases, the result of a natural process due to the exposure of the stain samples to the environment. Experiences with degraded DNA from casework samples show that every sample may exhibit different properties in this respect, and that it is difficult to systematically assess the performance of routinely used typing systems for the analysis of degraded DNA samples. Using a batch of artificially degraded DNA with an average fragment size of approx. 200 bp a collaborative exercise was carried out among 38 forensic laboratories from 17 European countries. The results were assessed according to correct allele detection, peak height and balance as well as the occurrence of artefacts. A number of common problems were identified based on these results such as strong peak imbalance in heterozygous genotypes for the larger short tandem repeat (STR) fragments after increased PCR cycle numbers, artefact signals and allelic drop-out. Based on the observations, strategies are discussed to overcome these problems. The strategies include careful balancing of the amount of template DNA and the PCR cycle numbers, the reaction volume and the amount of Taq polymerase. Furthermore, a careful evaluation of the results of the fragment analysis and of automated allele calling is necessary to identify the correct alleles and avoid artefacts.
Alleles↗
Encoded evidence: DNA in forensic analysis.
Sherlock Holmes said "it has long been an axiom of mine that the little things are infinitely the most important", but never imagined that such a little thing, the DNA molecule, could become perhaps the most powerful single tool in the multifaceted fight against crime. Twenty years after the development of DNA fingerprinting, forensic DNA analysis is key to the conviction or exoneration of suspects and the identification of victims of crimes, accidents and disasters, driving the development of innovative methods in molecular genetics, statistics and the use of massive intelligence databases.
DNA Fingerprinting↗
Development of a simulation model to assess the impact of contamination in casework using STRs.
Because contamination is usually tube-specific, negative controls cannot give assurance that an associated batch of extracted casework material is contaminant-free. However, it is possible to use them to predict the level of overall (undetected) contamination that is processed by an operational DNA unit. A MATLAB-based program was used to combine results of negative controls with actual casework DNA profiles to assess the probability that laboratory contaminants will give rise to reportable profiles (along with their likelihood ratios). Using data from an operational DNA unit as an example, it was demonstrated that the risk is inextricably linked to guidelines used to interpret DNA profiles. We have demonstrated how computer-based models can predict the levels of contamination expected in the process and, in addition, how the process can be made more robust by changing reporting guidelines. There is a need to compare DNA profiles against staff and plasticware elimination databases in order to determine sources of contamination. The likeliest outcome of a contamination event is false exclusion.
Computer Simulation↗
A comparison of adjustment methods to test the robustness of an STR DNA database comprised of 24 European populations.
An aim of the European Network of Forensic Science Institutes (ENFSI) is to produce a DNA database of second generation multiplex (SGM) STR profiles that is representative of the resident cosmopolitan populations. To achieve this, data were collected from 24 different populations. All of the data were combined to form one database of 5700 profiles from which allele proportions were calculated. The robustness of this combined European database was tested by estimating parameter d for every DNA profile, where d=log(10)(Pm(c)/Pm(E)) Pm(c) is the match probability of the profile calculated from its cognate database and Pm(E) is the match probability of the combined European database. Overall there was a small tendency for Pm(c)>Pm(E) primarily because of sampling bias. This bias was removed by the simple expediency of applying an adjustment factor to the calculation of Pm(E). These were selected from the Balding size bias correction, the Balding and Nichols Fst correction, a minimum allele proportion (between 0.01 and 0.02), an upper bound of a 95% confidence interval (CI) and a lower bound on the genotype match probability. It was demonstrated that a single European database is a feasible proposition. A combination of different adjustment methods can be used to ensure that the result is conservative relative to the cognate database, and their effect measured by parameter d.
Alleles↗
The propensity of individuals to deposit DNA and secondary transfer of low level DNA from individuals to inert surfaces.
We have shown that there is a difference between individuals in their tendency to deposit DNA on an item when it is touched. While a good DNA shedder may leave behind a full DNA profile immediately after hand washing, poor DNA shedders may only do so when their hands have not been washed for a period of 6h. We have also demonstrated that transfer of DNA from one individual (A) to another (B) and subsequently to an object is possible under specific laboratory conditions using the AMPFISTR SGM Plus multiplex at both 28 and 34 PCR cycles. This is a form of secondary transfer. If a 30 min or 1h delay was introduced before contact of individual B with the object then at 34 cycles a mixture of profiles from both individuals was recovered. We have also determined that the quantity and quality of DNA profiles recovered is dependent upon the particular individuals involved in the transfer process. The findings reported here are preliminary and further investigations are underway in order to further add to understanding of the issues of DNA transfer and persistence.
Alleles↗
Work rehabilitation and patterns of substance use among persons with schizophrenia.
OBJECTIVE: The aim of this study was to identify patterns of substance use among participants in work rehabilitation, to identify symptom patterns associated with substance use, and to assess the impact of substance use on work rehabilitation outcomes. METHODS: Addiction Severity Index interviews were conducted and Positive and Negative Syndrome Scale ratings were obtained at the start of the study and again five months and 12 months later for 220 patients with schizophrenia or schizoaffective disorder who were already enrolled in a study of work rehabilitation. RESULTS: Eighty percent of the participants met Addiction Severity Index criteria for lifetime substance use, but 75 percent were abstinent at intake. During the 12-month follow-up period, abstinence rates remained above 66 percent. Participants with a lifetime history of cocaine use were more likely to return to substance use. The type of substance used was also related to distinct symptom patterns. Participants with past cocaine use had more severe hostility symptoms and less severe negative symptoms. No significant relationships were found between lifetime or current substance use and participation in the work rehabilitation program. CONCLUSIONS: Even though a high rate of lifetime substance use was observed in this sample, most participants were in stable remission of substance use throughout the one-year study. Substance use and work participation appeared to be semiautonomous: substance use did not directly affect work participation, and work participation did not directly affect substance use.
Adult↗
Role of short tandem repeat DNA in forensic casework in the UK--past, present, and future perspectives.
The analysis of short tandem repeat (STR) DNA sequences is of fundamental importance to forensic science because they have become the recognized standard in constructing national public databases. Consequently, considerable effort has been expended in developing multiplexed (one tube) reactions that analyze several loci in combination. The implementation of STRs in casework cannot take place without a full understanding of the systems used. The purpose of validation is to characterize multiplexes when one is challenged with forensic samples. For example, mixtures are often encountered that may be particularly difficult to interpret against a background of allelic artifacts. By increasing the number of PCR amplification cycles, it is possible to dramatically boost the sensitivity of the system so that just a handful of cells may be successfully analyzed. However, interpretation is much more complex because the origin of DNA profiles may be less certain and complicated by issues such as contamination, the potential for innocent transfer and a predominance of mixtures. This review provides a brief historical background of the development of STRs in forensic casework that culminated in the creation of national DNA databases. The development of guidelines to interpret complex DNA profiles, such as mixtures, is outlined. Finally, the recent innovation of low copy number DNA profiling is explained along with the special considerations needed to report in court.
DNA↗
An assessment of whether SNPs will replace STRs in national DNA databases--joint considerations of the DNA working group of the European Network of Forensic Science Institutes (ENFSI) and the Scientific Working Group on DNA Analysis Methods (SWGDAM).
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DNA Fingerprinting↗