Institute of Legal Medicine, Catholic University of Sacred Heart, School of Medicine, Largo Francesco Vito 1, 00168 Rome, Italy.
Forensic Sci Int Genet. 2012 Dec;6(6):735-48. doi: 10.1016/j.fsigen.2012.08.005. Epub 2012 Sep 1.
Evaluation of series of PCR experiments referring to the same evidence is not infrequent in a forensic casework. This situation is met when 'series of results in mixture' (EPGs produced by reiterating PCR experiments over the same DNA mixture extract) have to be interpreted or when 'potentially related traces' (mixtures that can have contributors in common) require a combined interpretation. In these cases, there can be uncertainty on the genotype assignment, since: (a) more than one genotype combination fall under the same peak profile; (b) PCR preferential amplification alters pre-PCR allelic proportions; (c) other, more unpredictable technical problems (dropouts/dropins, etc.) take place. The uncertainty in the genotype assignment is in most cases addressed by empirical methods (selection of just one particular profile; extraction of consensual or composite profiles) that disregard part of the evidence. Genotype assignment should conversely take advantage from a joint Bayesian analysis (JBA) of all STRs peak areas generated at each experiment. This is the typical case of Bayesian analysis in which adoption of object-oriented Bayesian networks (OOBNs) could be highly helpful. Starting from experimentally designed mixtures, we created typical examples of 'series of results in mixture' of 'potentially related traces'. JBA was some administered to the whole peak area evidence, by specifically tailored OOBNs models, which enabled genotype assignment reflecting all the available evidence. Examples of a residual ambiguity in the genotype assignment came to light at assumed genotypes with partially overlapping alleles (for example: AB+AC→ABC). In the 'series of results in mixture', this uncertainty was in part refractory to the joint evaluation. Ambiguity was conversely dissipated at the 'potentially related' trace example, where the ABC allelic scheme at the first trace was interpreted together with other unambiguous combinations (ABCD; AB) at the related trace. We emphasize the need to carry out extensive, blind sensitivity tests specifically addressing the residual ambiguity that arises from overlapping results mixed at various quantitative ratios.
在法医学案件中,经常需要评估涉及同一证据的一系列 PCR 实验。这种情况出现在需要解释“混合物中的系列结果”(通过重复 PCR 实验在同一 DNA 混合物提取物上产生的 EPG)或需要联合解释“潜在相关痕迹”(可以有共同贡献者的混合物)时。在这些情况下,基因型赋值可能存在不确定性,因为:(a) 多个基因型组合符合相同的峰型;(b) PCR 优先扩增改变了预 PCR 等位基因比例;(c) 发生其他更不可预测的技术问题(缺失/插入等)。基因型赋值的不确定性在大多数情况下通过经验方法(选择一个特定的图谱;提取共识或复合图谱)来解决,这些方法忽略了部分证据。相反,基因型赋值应该利用所有 STR 峰面积的联合贝叶斯分析(JBA),每个实验都生成这些峰面积。这是贝叶斯分析的典型情况,采用面向对象的贝叶斯网络(OOBN)可能会非常有帮助。从实验设计的混合物开始,我们创建了“潜在相关痕迹”的“混合物中的系列结果”的典型示例。通过专门设计的 OOBNs 模型,对整个峰面积证据进行 JBA,从而反映所有可用证据的基因型赋值。在假设的基因型部分重叠等位基因的情况下(例如:AB+AC→ABC),基因型赋值出现残余不确定性的例子。在“潜在相关”痕迹的例子中,这种不确定性在一定程度上可以通过联合评估来解决。相反,在 ABC 等位基因方案在第一个痕迹上与相关痕迹上其他明确的组合(ABCD;AB)一起解释时,不确定性得到了消除。我们强调需要进行广泛的、盲目的敏感性测试,专门解决由于以各种定量比例混合而产生的剩余不确定性。