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使用 Rapid DNA 仪器处理模拟放射性恐怖事件中的生物样本。

Processing biological samples from simulated radiological terrorist events using Rapid DNA instruments.

机构信息

Royal Canadian Mounted Police, Forensic Science & Identification Services, National Forensic Laboratory Services, Biology Policy & Program Support, 1200 Vanier Parkway, Ottawa, Ontario K1A 0R2, Canada.

Royal Canadian Mounted Police, Forensic Science & Identification Services, Science & Strategic Policy, 1200 Vanier Parkway, Ottawa, Ontario K1A 0R2, Canada.

出版信息

Forensic Sci Int. 2024 Jan;354:111887. doi: 10.1016/j.forsciint.2023.111887. Epub 2023 Nov 19.

Abstract

Two commercially available portable Rapid DNA instruments were evaluated for their ability to process 1 µL and 10 µL saliva samples deposited on metal and plastic surfaces and contaminated with surrogates of cesium (Cs)-137, strontium (Sr)-90 and cobalt (Co)-60; radioactive materials potentially released during a nuclear weapon accident or a radiological dispersal device detonation. A comparable success rate was noted for both Rapid DNA instruments when considering the number of complete and balanced DNA profiles, the number of profiles with a minimum of 10 autosomal STR loci (out of 23 [FlexPlex™ 27] or 21 [GlobalFiler™ Express]), and the possibility to search a national DNA database in Canada and the United States. Cobalt had an adverse impact on the quality of the megaplex short tandem repeat (STR) DNA profiles derived on each instrument for two of the three contamination levels tested in this study, i.e., 0.05 M and 0.1 M as reflected by a reduced number of detected alleles and decreased profile peak heights. Strontium exhibited some adverse effect on the Rapid DNA results when used at the highest contamination level (0.1 M) whereas cesium had none. No new artifacts were observed in the Rapid DNA profiles of samples spiked with the non-radiogenic surrogates. Importantly, in the context of a radiological/nuclear (RN) event, the ANDE™ 6C offers the possibility to dispose of all radioactive materials associated with contaminated samples quickly using a chip on which all steps of the Rapid DNA process are performed whereas the RapidHIT™ ID accumulates radioactive materials for many days before disposal. An individual handling 25 samples in a week (5 per day) on the RapidHIT™ ID at a 30.5 cm distance with a 5 min exposure to the radioactive source estimated at every run would exceed the 0.042 µSv/5 min limit with gamma dose rates for Cs at 0.13 mSv and for Co at 3.8 mSv. Beta dose rates calculated for the surrogate isotopes at the three concentrations tested were also above the recommended radiation exposure limit of 1 mSv/yr (0.042 µSv/5 min). Various potential mechanisms of action behind the interference noted for Sr and Co at high concentrations are presented. These elements may play a role in the steps prior to PCR (at the DNA molecule by binding to bases or to phosphate groups), during PCR (at the DNA polymerase as cofactors for catalytic sites), or even during amplified DNA fragment detection (as fluorescence quenchers).

摘要

两种市售的便携式 Rapid DNA 仪器被评估用于处理分别沉积在金属和塑料表面上的 1 µL 和 10 µL 唾液样本,这些样本受到铯 (Cs)-137、锶 (Sr)-90 和钴 (Co)-60 替代物的污染;这些放射性材料可能是在核武器事故或放射性分散装置爆炸期间释放的。考虑到完整和平衡的 DNA 谱的数量、具有至少 10 个常染色体 STR 基因座(23 个 [FlexPlex™ 27] 或 21 个 [GlobalFiler™ Express])的谱的数量,以及在加拿大和美国搜索国家 DNA 数据库的可能性,两种 Rapid DNA 仪器的成功率相当。钴对两种污染水平(本研究测试的 0.05 M 和 0.1 M)下从每个仪器获得的 megaplex 短串联重复 (STR) DNA 谱的质量有不利影响,表现为检测到的等位基因数量减少和谱峰高度降低。当在最高污染水平(0.1 M)下使用锶时,Rapid DNA 结果显示出一些不利影响,而铯则没有。在添加非放射性替代物的样品的 Rapid DNA 谱中没有观察到新的伪影。重要的是,在放射性/核(RN)事件的情况下,ANDE™ 6C 提供了一种可能性,可以使用快速处理所有与污染样品相关的放射性材料芯片,而 RapidHIT™ ID 在处理之前会积累数天的放射性材料放射性物质。一个人每周在 RapidHIT™ ID 上处理 25 个样本(每天 5 个),与放射性源的距离为 30.5 厘米,每次运行时暴露 5 分钟,估计 γ 剂量率对于 Cs 为 0.13 mSv,对于 Co 为 3.8 mSv,将超过 0.042 µSv/5 min 限值。对于测试的三种浓度的替代同位素,计算出的 β 剂量率也超过了 1 mSv/yr(0.042 µSv/5 min)的推荐辐射暴露限值。还提出了高浓度下 Sr 和 Co 干扰的各种潜在作用机制。这些元素可能在 PCR 之前的步骤中发挥作用(在 DNA 分子上通过与碱基或磷酸基团结合)、PCR 期间(在 DNA 聚合酶上作为催化位点的辅助因子),甚至在扩增的 DNA 片段检测期间(作为荧光猝灭剂)。

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