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NucleoSpin®法医滤器对痕量 DNA 拭子中 DNA 回收的影响。

The effect of NucleoSpin® Forensic Filters on DNA recovery from trace DNA swabs.

机构信息

Division of Forensic Medicine and Toxicology, Department of Pathology, Faculty of Health Science, University of Cape Town, South Africa.

Division of Forensic Medicine and Toxicology, Department of Pathology, Faculty of Health Science, University of Cape Town, South Africa.

出版信息

Sci Justice. 2022 May;62(3):284-287. doi: 10.1016/j.scijus.2022.03.001. Epub 2022 Mar 8.

Abstract

Forensic DNA profiling is a globally accepted method for human identification, however, obtaining full DNA profiles from trace DNA can be challenging. The optimal recovery of DNA from trace DNA swabs is therefore crucial. Methods for extracting DNA from swabs often make use of a spin basket combined with a centrifugation step, to enhance the release of cells from the swab prior to DNA extraction. The NucleoSpin® Forensic Filter (Macherey-Nagel, Düren) is a type of spin basket, but it has not been thoroughly assessed on trace DNA samples. This study aimed to assess if the inclusion of the NucleoSpin® Forensic Filter significantly improved DNA recovery and DNA profiling success from cotton and flocked swabs used to collect trace DNA and buccal cells (control). Buccal cells and trace DNA samples were collected from 25 volunteers using each swab type (cotton and flocked) in duplicate. DNA was extracted from the samples using the NucleoSpin® DNA Forensic kit, one set with, and the other set without, NucleoSpin® Forensic Filters. DNA concentration was assessed using real time PCR, and DNA profiling was done using the PowerPlex® ESX 16 system. The inclusion of the NucleoSpin® Forensic Filters significantly improved DNA concentration for buccal cells that were collected using flocked swabs (p = 0.035). However, no significant differences were noted for trace DNA samples for either swab type. There was also no significant difference in DNA profiling success when NucleoSpin® Forensic Filters were used, regardless of swab and sample type. These results may be helpful for laboratories that are considering the NucleoSpin® Forensic Filters in the DNA extraction workflow, particularly for trace DNA samples.

摘要

法医 DNA 分析是一种全球公认的人类身份识别方法,然而,从痕量 DNA 中获取完整的 DNA 谱可能具有挑战性。因此,从痕量 DNA 拭子中最佳地回收 DNA 至关重要。从拭子中提取 DNA 的方法通常利用带有离心步骤的旋转篮,以增强细胞从拭子中释放出来,然后再进行 DNA 提取。NucleoSpin®法医过滤器(美沙东-内盖尔,杜伦)是一种旋转篮,但尚未对痕量 DNA 样本进行彻底评估。本研究旨在评估是否包括 NucleoSpin®法医过滤器可以显著提高从用于收集痕量 DNA 和口腔细胞(对照)的棉拭子和植绒拭子中回收 DNA 和 DNA 图谱构建的成功率。使用每种拭子类型(棉拭子和植绒拭子)从 25 名志愿者中采集口腔细胞和痕量 DNA 样本,每个样本一式两份。使用 NucleoSpin®DNA 法医试剂盒从样本中提取 DNA,一组包含,另一组不包含 NucleoSpin®法医过滤器。使用实时 PCR 评估 DNA 浓度,使用 PowerPlex®ESX 16 系统进行 DNA 图谱构建。在使用植绒拭子采集的口腔细胞中,包含 NucleoSpin®法医过滤器可显著提高 DNA 浓度(p=0.035)。然而,对于两种拭子类型的痕量 DNA 样本,均未观察到显著差异。使用 NucleoSpin®法医过滤器时,无论拭子和样本类型如何,DNA 图谱构建的成功率也没有显著差异。这些结果可能有助于正在考虑在 DNA 提取工作流程中使用 NucleoSpin®法医过滤器的实验室,特别是对于痕量 DNA 样本。

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