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改良对二甲氨基肉桂醛溶液的研制及其在 STR 分析中的应用。

Development of a modified p-dimethylaminocinnamaldehyde solution for touch DNA analysis and its application to STR analysis.

机构信息

Forensic Science Laboratory, Hyogo Prefectural Police Headquarters, 5-4-1 Shimoyamate-dori, Chuo-ku, Kobe 650-8510, Japan.

Forensic Science Laboratory, Hyogo Prefectural Police Headquarters, 5-4-1 Shimoyamate-dori, Chuo-ku, Kobe 650-8510, Japan.

出版信息

Forensic Sci Int Genet. 2019 Jan;38:86-92. doi: 10.1016/j.fsigen.2018.10.007. Epub 2018 Oct 12.

Abstract

We developed an acid-free p-dimethylaminocinnamaldehyde (DMAC) solution containing silicone oil that was suitable for spraying on clothing for analysis of biological samples such as touch DNA. We investigated the effect of this solution and irradiation with blue light emitting diode (LED) light on short tandem repeat (STR) analysis. To examine the effect of adding acid to the DMAC solution on visualizing biological samples, saliva sample was deposited on T-shirt. The T-shirt was sprayed with acid-added DMAC or acid-free DMAC solution and left for 2 h before irradiation with the blue LED light. We observed no differences between the fluorescence intensities achieved with these two solutions. To examine the effect of acid addition to the DMAC solution on STR analysis, sweat samples were smeared on glass slides and dried. The slides were sprayed with acid-added or acid-free DMAC solution and irradiated with the blue LED light. Samples were collected from the slides with swabs, and DNA was extracted from each sample using a PrepFiler Express™ Forensic DNA Extraction Kit and quantified using a Human DNA quantification kit (Takara RR281). The extracted DNA was amplified using an AmpFℓSTR Identifiler Plus PCR Amplification Kit for STR typing. We found that addition of acid to DMAC had little effect on DNA contained in the biological samples and STR analysis. To investigate whether DMAC could be used to visualize biological samples on clothes, saliva, sweat, and finger and palm prints were deposited on separate T-shirts. Biological samples were treated with DMAC and observed after 2 h, 1, 2, or 3 days under the blue LED. All biological samples were visualized and emitted fluorescence after 2 h. To examine the effects of the DMAC solution and LED irradiation on STR analysis, including DNA extraction, quantification, and STR typing, saliva and sweat were smeared on glass slides and dried. Touch DNA samples were deposited on glass slides directly. The slides were then sprayed with DMAC solution, sprayed with DMAC solution and irradiated with the blue LED, or left untreated. Samples were collected from the slides with swabs, and DNA was extracted, quantified, and amplified using the above kits. These results suggest that the DMAC solution and blue LED light will have no adverse effects on STR analysis. Therefore, this method will be very useful for touch DNA analysis in forensic investigations.

摘要

我们开发了一种不含酸的 p-二甲基氨肉桂醛 (DMAC) 溶液,其中含有硅油,适用于喷涂在衣物上,用于分析接触 DNA 等生物样本。我们研究了该溶液和蓝光发光二极管 (LED) 光照射对短串联重复序列 (STR) 分析的影响。为了研究在 DMAC 溶液中添加酸对生物样本可视化的影响,我们将唾液样本沉积在 T 恤上。用添加酸的 DMAC 或无酸 DMAC 溶液喷涂 T 恤,然后在蓝光 LED 光下照射 2 小时。我们观察到这两种溶液的荧光强度没有差异。为了研究在 DMAC 溶液中添加酸对 STR 分析的影响,我们将汗渍涂抹在载玻片上并使其干燥。用添加酸的或无酸 DMAC 溶液喷涂载玻片,然后用蓝光 LED 光照射。用棉签从载玻片上收集样本,然后使用 PrepFiler Express™法医 DNA 提取试剂盒从每个样本中提取 DNA,并使用 Human DNA quantification kit(Takara RR281)定量。使用 AmpFℓSTR Identifiler Plus PCR 扩增试剂盒对提取的 DNA 进行 STR 分型。我们发现,在 DMAC 中添加酸对生物样本中的 DNA 几乎没有影响,对 STR 分析也没有影响。为了研究 DMAC 是否可用于在衣物上可视化生物样本,我们将唾液、汗液、手指和手掌印分别沉积在不同的 T 恤上。用 DMAC 处理生物样本,然后在蓝光 LED 下放置 2 小时、1 天、2 天或 3 天进行观察。所有生物样本在 2 小时后均可见并发出荧光。为了研究 DMAC 溶液和 LED 照射对 STR 分析的影响,包括 DNA 提取、定量和 STR 分型,我们将唾液和汗液涂抹在载玻片上并使其干燥。直接将接触 DNA 样本沉积在载玻片上。然后用 DMAC 溶液喷涂载玻片,或用 DMAC 溶液喷涂后用蓝光 LED 照射,或不处理。用棉签从载玻片上收集样本,使用上述试剂盒提取、定量和扩增 DNA。这些结果表明,DMAC 溶液和蓝光 LED 光不会对 STR 分析产生不利影响。因此,该方法将非常有助于法医调查中的接触 DNA 分析。

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