Development of a modified p-dimethylaminocinnamaldehyde solution for touch DNA analysis and its application to STR analysis
Development of a modified p-dimethylaminocinnamaldehyde solution for touch DNA analysis and its application to STR analysis
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用于接触 DNA 分析的改良对二甲氨基肉桂醛溶液的开发及其在 STR 分析中的应用
DOI：
10.1016/j.fsigen.2018.10.007
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发表时间：
2019
影响因子：
2.2
通讯作者：
Ryohei Yano. Osamu Shimoda. Takaaki Okitsu. Makoto Sakurada. Yasuhiro Ueno
中科院分区：
文献类型：
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作者：
助永壮平;田代公則;柴田浩幸;Ryohei Yano. Osamu Shimoda. Takaaki Okitsu. Makoto Sakurada. Yasuhiro Ueno
我们开发了一种含硅油的无酸二甲基氨基肉桂醛（DMAC）溶液，适用于喷涂在衣服上，用于分析触摸DNA等生物样品。我们研究了该溶液和蓝光发光二极管（LED）灯照射对短串联重复序列（STR）分析的影响。为了检查向DMAC溶液中加入酸对可视化生物样品的影响，将唾液样品沉积在T恤上。用添加酸的DMAC或无酸的DMAC溶液喷涂T恤，并放置2小时，然后用蓝色LED灯照射。我们观察到这两种溶液所获得的荧光强度之间没有差异。为了检查向DMAC溶液中添加酸对STR分析的影响，将汗液样品涂抹在载玻片上并干燥。用加酸或无酸的DMAC溶液喷洒载玻片，并用蓝色LED灯照射。用拭子从载玻片收集样品，并且使用PrepFiler Express™法医DNA提取试剂盒从每个样品提取DNA，并且使用人DNA定量试剂盒（Takara RR 281）定量。提取的DNA使用用于STR分型的AmpF STR®Identifiler®Plus PCR扩增试剂盒进行扩增。我们发现，在DMAC中加入酸对生物样品中所含的DNA和STR分析几乎没有影响。为了研究DMAC是否可以用于可视化衣服上的生物样本，唾液，汗液，手指和手掌印被沉积在单独的T恤上。用DMAC处理生物样品，并在2小时、1天、2天或3天后在蓝色LED下观察。所有生物样品在2小时后可视化并发射荧光。为了检查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，将唾液和汗液涂抹在载玻片上并干燥。Touch DNA样品直接沉积在载玻片上。然后将载玻片用DMAC溶液喷雾，用DMAC溶液喷雾并用蓝色LED照射，或不进行处理。用拭子从载玻片中收集样本，并使用上述试剂盒提取、定量和扩增DNA。这些结果表明，DMAC溶液和蓝色LED灯不会对STR分析产生不利影响。因此，该方法将是非常有用的接触DNA分析在法医学调查。
We developed an acid-freep-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.