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      Verifying likelihoods for low template DNA profiles using multiple replicates

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          Highlights

          • The behaviour of multi-replicate LRs with respect to the inverse match probability is proposed as a method to validate forensic LR software.

          • We perform lab-based and simulated experiments of one-, two- and three-contributor CSPs, as well as investigating a real-world CSP.

          • LRs rise towards the IMP with additional replicates, while never exceeding it. Additionally, the LR from multiple low-template replicates can exceed that from a single good-quality sample.

          • We validate likeLTD by demonstrating that it adheres to the expected behaviours.

          Abstract

          To date there is no generally accepted method to test the validity of algorithms used to compute likelihood ratios (LR) evaluating forensic DNA profiles from low-template and/or degraded samples. An upper bound on the LR is provided by the inverse of the match probability, which is the usual measure of weight of evidence for standard DNA profiles not subject to the stochastic effects that are the hallmark of low-template profiles. However, even for low-template profiles the LR in favour of a true prosecution hypothesis should approach this bound as the number of profiling replicates increases, provided that the queried contributor is the major contributor. Moreover, for sufficiently many replicates the standard LR for mixtures is often surpassed by the low-template LR. It follows that multiple LTDNA replicates can provide stronger evidence for a contributor to a mixture than a standard analysis of a good-quality profile. Here, we examine the performance of the likeLTD software for up to eight replicate profiling runs. We consider simulated and laboratory-generated replicates as well as resampling replicates from a real crime case. We show that LRs generated by likeLTD usually do exceed the mixture LR given sufficient replicates, are bounded above by the inverse match probability and do approach this bound closely when this is expected. We also show good performance of likeLTD even when a large majority of alleles are designated as uncertain, and suggest that there can be advantages to using different profiling sensitivities for different replicates. Overall, our results support both the validity of the underlying mathematical model and its correct implementation in the likeLTD software.

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          Most cited references22

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          DNA commission of the International Society of Forensic Genetics: Recommendations on the interpretation of mixtures.

          The DNA commission of the International Society of Forensic Genetics (ISFG) was convened at the 21st congress of the International Society for Forensic Genetics held between 13 and 17 September in the Azores, Portugal. The purpose of the group was to agree on guidelines to encourage best practice that can be universally applied to assist with mixture interpretation. In addition the commission was tasked to provide guidance on low copy number (LCN) reporting. Our discussions have highlighted a significant need for continuing education and research into this area. We have attempted to present a consensus from experts but to be practical we do not claim to have conveyed a clear vision in every respect in this difficult subject. For this reason, we propose to allow a period of time for feedback and reflection by the scientific community. Then the DNA commission will meet again to consider further recommendations.
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            Validity of low copy number typing and applications to forensic science.

            Low copy number (LCN) typing, particularly for current short tandem repeat (STR) typing, refers to the analysis of any sample that contains less than 200 pg of template DNA. Generally, LCN typing simply can be defined as the analysis of any DNA sample where the results are below the stochastic threshold for reliable interpretation. There are a number of methodologies to increase sensitivity of detection to enable LCN typing. These approaches encompass modifications during the polymerase chain reaction (PCR) and/or post-PCR manipulations. Regardless of the manipulations, when processing a small number of starting templates during the PCR exaggerated stochastic sampling effects will occur. The result is that several phenomena can occur: a substantial imbalance of 2 alleles at a given heterozygous locus, allelic dropout, or increased stutter. With increased sensitivity of detection there is a concomitant increased risk of contamination. Recently, a commission reviewed LCN typing and found it to be "robust" and "fit for purpose." Because LCN analysis by its nature is not reproducible, it cannot be considered as robust as that associated with conventional DNA typing. The findings of the commission seem inconsistent with the nature of LCN typing. While LCN typing is appropriate for identification of missing persons and human remains and for developing investigative leads, caution should be taken with its use in other endeavors until developments are made that overcome the vagaries of LCN typing. A more in-depth evaluation by the greater scientific community is warranted. The issues to consider include: training and education, evidence handling and collection procedures, the application or purpose for which the LCN result will be used, the reliability of current LCN methods, replicate analyses, interpretation and uncertainty, report writing, validation requirements, and alternate methodologies for better performance.
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              Interpreting low template DNA profiles.

              We discuss the interpretation of DNA profiles obtained from low template DNA samples. The most important challenge to interpretation in this setting arises when either or both of "drop-out" and "drop-in" create discordances between the crime scene DNA profile and the DNA profile expected under the prosecution allegation. Stutter and unbalanced peak heights are also problematic, in addition to the effects of masking from the profile of a known contributor. We outline a framework for assessing such evidence, based on likelihood ratios that involve drop-out and drop-in probabilities, and apply it to two casework examples. Our framework extends previous work, including new approaches to modelling homozygote drop-out and uncertainty in allele calls for stutter, masking and near-threshold peaks. We show that some current approaches to interpretation, such as ignoring a discrepant locus or reporting a "Random Man Not Excluded" (RMNE) probability, can be systematically unfair to defendants, sometimes extremely so. We also show that the LR can depend strongly on the assumed value for the drop-out probability, and there is typically no approximation that is useful for all values. We illustrate that ignoring the possibility of drop-in is usually unfair to defendants, and argue that under circumstances in which the prosecution relies on drop-out, it may be unsatisfactory to ignore any possibility of drop-in.
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                Author and article information

                Contributors
                Journal
                Forensic Sci Int Genet
                Forensic Sci Int Genet
                Forensic Science International. Genetics
                Elsevier
                1872-4973
                1878-0326
                1 November 2014
                November 2014
                : 13
                : 82-89
                Affiliations
                [a ]UCL Genetics Institute, Darwin Building, Gower Street, London WC1E 6BT, UK
                [b ]Orchid Cellmark Ltd., Abingdon Business Park, Blacklands Way, Abingdon OX14 1YX, UK
                Author notes
                [* ]Corresponding author. c.steele.11@ 123456ucl.ac.uk
                Article
                S1872-4973(14)00136-7
                10.1016/j.fsigen.2014.06.018
                4234080
                25082140
                4ceafdf8-c4d2-4b51-aa6f-ca6b4fe06299
                © 2014 The Authors
                History
                : 19 February 2014
                : 9 June 2014
                : 30 June 2014
                Categories
                Article

                Forensic science
                low-template dna,dna mixtures,likelihood ratio,replicates,forensic,likeltd
                Forensic science
                low-template dna, dna mixtures, likelihood ratio, replicates, forensic, likeltd

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