Abstract：
Over a decade after the publication of the first forensic DNA phenotyping (FDP) studies, DNA-based appearance predictions are now becoming a reality in routine crime scene investigations. The significant number of publications dedicated to the subject of FDP clearly demonstrates a sustained interest and a strong need for further method development. However, the implementation of FDP in routine work still encounters obstacles, and one of these challenges is making phenotype predictions from DNA mixtures. In this study, we examined single-cell sequencing as a potential tool to enable reliable phenotyping of contributors within mixtures. Two mock mixtures, each containing two contributors with similar and different physical appearances, were analyzed using two different workflows. In the first workflow, the mixtures were sequenced using the Ion AmpliSeq™ PhenoTrivium Panel, which includes 41 HIrisPlex-S (HPS) markers. Subsequently, the genotypes were analyzed using the HPS Deconvolution Tool to predict the phenotypes of both contributors. The second workflow involved the introduction of single-cell separation and collection using the DEPArray™ PLUS System. Two different PhenoTrivium amplification protocols were tested, and the phenotype predictions from single cells were compared with the results obtained using the HPS Tool. Our results suggest that the approach presented here allows for the obtainment of nearly complete HIrisPlex-S profiles with accurate genotypes and reliable phenotype predictions from single cells. This method proves successful in deconvoluting mixtures submitted to forensic DNA phenotyping.
摘要：
在第一个法医DNA表型（FDP）研究发表十年后，基于DNA的外观预测现在在常规犯罪现场调查中已成为现实。致力于FDP主题的大量出版物清楚地表明了对进一步方法开发的持续兴趣和强烈需求。然而,FDP在日常工作中的实施仍然遇到障碍，这些挑战之一是从DNA混合物中进行表型预测。在这项研究中,我们研究了单细胞测序作为一种潜在工具,可以对混合物中的贡献者进行可靠的表型分析.两种模拟混合物，每个包含两个具有相似和不同物理外观的贡献者，使用两个不同的工作流程进行了分析。在第一个工作流程中，使用IonAmpliSeq™PhenoTrivium面板对混合物进行测序，其中包括41个HIrisPlex-S（HPS）标记。随后，使用HPS解卷积工具分析基因型,以预测两个贡献者的表型.第二个工作流程涉及使用DEPArray™PLUS系统引入单细胞分离和收集。测试了两种不同的PhenoTrivium扩增方案，并且将来自单细胞的表型预测与使用HPS工具获得的结果进行比较。我们的结果表明，这里提出的方法可以获得几乎完整的HIrisPlex-S谱，具有准确的基因型和可靠的单细胞表型预测。该方法在对提交给法医DNA表型的混合物进行解卷积方面被证明是成功的。
