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评估 Diamond™ 核酸染料在法医学实际工作中用于潜伏 DNA 鉴定和靶向取样。

Assessment of Diamond™ Nucleic Acid Dye for the identification and targeted sampling of latent DNA in operational casework.

机构信息

Forensic Science SA, GPO Box 2790, Adelaide, South Australia 5001, Australia.

Forensic Science SA, GPO Box 2790, Adelaide, South Australia 5001, Australia; College of Science and Engineering, Flinders University of South Australia, Bedford Park, South Australia 5042, Australia.

出版信息

Forensic Sci Int Genet. 2021 Nov;55:102579. doi: 10.1016/j.fsigen.2021.102579. Epub 2021 Aug 21.

Abstract

Recovery and DNA profiling of latent touch DNA deposits is a ubiquitous practice by operational forensic laboratories that provides critical evidence in many criminal investigations. Despite recent improvements in the sensitivity of contemporary DNA profiling kits, the inability to localise and visually quantify touch DNA deposits on an exhibit means that ineffective or unwarranted sampling is often performed leading to poor success. Diamond™ Nucleic Acid Dye (DD) is a fluorescent DNA binding dye which has recently been shown to bind to corneocytes enabling visualisation and targeted sampling of touch DNA deposits under controlled conditions. The ability to translate these findings to operational casework, where a diverse range of substrates is encountered and the amount and distribution of touch DNA is uncontrolled, is currently unknown. Here, we provide the first report on the use of DD in an operational context. Spraying items with DD was shown to have no impact on downstream immunological testing, DNA extraction, or DNA profiling with the GlobalFiler™ PCR amplification kit. DD was shown to effectively locate areas of touch DNA on select exhibits using the Polilight. Issues with background fluorescence, non-specific staining, interference from fingerprint enhancement reagents, or absorbance of the excitation light by black surfaces demonstrated that DD is not compatible with all exhibits. Background fluorescence also prevented the use of DD to screen for the presence of cellular material on IsoHelix swabs post-sampling but it was suitable for screening Lovell DNA tapelifts. A casework trial of 49 plastic bag and tape exhibits showed limited application of DD to triage out negative items as DNA was recovered from items where DD fluorescence was not detected. Where DD fluorescence was detected, its broad distribution prevented targeted sampling and any correlation to be made between the amount observed and DNA yield or profiling outcome. The DD procedure also increased the time taken to search exhibits and risk of inadvertent contamination. Our study suggests that DD is not suited as a generalised screening technique across all touch casework exhibits but further investigation is warranted to determine its applicability to specific exhibit types.

摘要

恢复和 DNA 分析潜伏触摸 DNA 沉积物是操作法医实验室普遍采用的做法,可为许多刑事调查提供关键证据。尽管当代 DNA 分析试剂盒的灵敏度最近有所提高,但无法对展品上的触摸 DNA 沉积物进行定位和直观定量意味着,通常会进行无效或不必要的采样,从而导致成功率低下。钻石™核酸染料(DD)是一种荧光 DNA 结合染料,最近已被证明可与角质细胞结合,从而在受控条件下对触摸 DNA 沉积物进行可视化和靶向采样。将这些发现转化为操作案例的能力,在这些案例中,会遇到各种不同的基质,触摸 DNA 的数量和分布是不可控的,目前尚不清楚。在这里,我们首次报告了 DD 在操作环境中的使用。DD 喷雾对下游免疫检测、DNA 提取或使用 GlobalFiler™PCR 扩增试剂盒进行 DNA 分析均无影响。DD 被证明可以有效地在选定的展品上定位触摸 DNA 区域使用 Polilight。荧光背景、非特异性染色、指纹增强试剂的干扰或黑色表面对激发光的吸收等问题表明,DD 与所有展品均不兼容。背景荧光还阻止了使用 DD 来筛选采样后 IsoHelix 拭子上是否存在细胞物质,但它适合于筛选 Lovell DNA 胶带。对 49 个塑料袋和胶带展品进行的案例试验表明,DD 的应用有限,可用于对阴性物品进行分类,因为从没有检测到 DD 荧光的物品中回收了 DNA。在检测到 DD 荧光的地方,其广泛的分布阻止了靶向采样,并且无法观察到观察到的数量与 DNA 产量或分析结果之间的任何相关性。DD 程序还增加了搜索展品的时间和意外污染的风险。我们的研究表明,DD 不适合作为所有触摸案件展品的通用筛选技术,但需要进一步研究以确定其对特定展品类型的适用性。

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