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两种用于处理血液和唾液样本的 Rapid DNA 技术的比较分析。

Comparative analysis of two Rapid DNA technologies for the processing of blood and saliva-based samples.

机构信息

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

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

出版信息

Forensic Sci Int Genet. 2023 Nov;67:102928. doi: 10.1016/j.fsigen.2023.102928. Epub 2023 Aug 8.

Abstract

Rapid DNA technologies recently gained significant momentum as a means to generate DNA profiles faster than with standard laboratory workflows. Initially developed for the analysis of buccal reference samples, applications are being considered for other types of forensic samples. In this study, an identical set of 150 blood and saliva-based samples was processed using two different Rapid DNA technologies, the Applied BioSystems™ RapidHIT™ ID System using the RapidINTEL™ sample cartridge and the ANDE™ 6C Rapid DNA Analysis™ System using the I-Chip. A subset of samples were subjected to alternative collection methods or sample pre-treatments to determine the optimal strategy for each instrument. An equivalent sample set was also processed using a conventional DNA analysis workflow. The sensitivity range of the two Rapid DNA technologies was comparable based on blood and saliva dilution series, with both technologies able to generate full profiles from samples typically yielding 5-10 ng of DNA when processed using conventional DNA analysis. The brand of cotton swabs used for Rapid DNA analysis had an impact on the results for both systems. Differences were observed in success rate between the two systems when processing blood (on fabrics, FTA paper or hard surfaces) and saliva-based samples (drink containers, FTA paper, chewing gum, cigarette butt filter paper) and depended on the sample type. Importantly, deviating from the manufacturer's instructions for sample collection and pre-treatment was more detrimental to the ANDE 6C results. The quality of DNA profiles, as assessed using heterozygote peak height ratios, interloci balance and artifact presence, confirmed the results to be reliable and acceptable for single source samples. Profiling results were obtained when samples were reprocessed using the same Rapid DNA technology or conventional DNA analysis. Secondary analysis using a substitute software (GeneMapper ID-X v1.5) to recover additional genetic information was shown to be feasible. Finally, a comparison between the Applied Biosystems™ RapidHIT™ ID System Software v1.3.1 and v1.3.2 was also performed. Findings of this study could assist those interested in using Rapid DNA technology for blood or saliva-based samples, in various settings and for different applications.

摘要

快速 DNA 技术最近作为一种比标准实验室工作流程更快地生成 DNA 图谱的手段获得了显著的发展。最初开发用于分析口腔参考样本,现在正在考虑将其应用于其他类型的法医样本。在这项研究中,使用两种不同的快速 DNA 技术(使用 RapidINTEL 样品盒的 Applied BioSystems RapidHIT ID 系统和使用 I-Chip 的 ANDE 6C Rapid DNA Analysis System)处理了相同的 150 个血液和唾液样本。一部分样本采用了替代采集方法或样本预处理,以确定每种仪器的最佳策略。还使用传统 DNA 分析工作流程处理了一组等效样本。基于血液和唾液稀释系列,两种快速 DNA 技术的灵敏度范围相当,当使用传统 DNA 分析处理时,这两种技术都能够从通常产生 5-10ng DNA 的样本中生成完整图谱。用于快速 DNA 分析的棉签品牌对两种系统的结果都有影响。在处理血液(在织物、FTA 纸或硬表面上)和基于唾液的样本(饮料容器、FTA 纸、口香糖、香烟滤嘴过滤纸)时,两种系统之间的成功率存在差异,这取决于样本类型。重要的是,偏离制造商关于样本采集和预处理的说明对 ANDE 6C 的结果更为不利。使用杂合峰高比、基因座间平衡和伪影存在评估 DNA 谱的质量,证实结果可靠且可接受,适用于单一来源样本。使用相同的快速 DNA 技术或传统 DNA 分析重新处理样本时,即可获得分析结果。使用替代软件(GeneMapper ID-X v1.5)进行二次分析以恢复额外遗传信息是可行的。最后,还比较了 Applied Biosystems RapidHIT ID System Software v1.3.1 和 v1.3.2。本研究的结果可为有兴趣在各种环境和不同应用中使用基于血液或唾液的快速 DNA 技术的人提供帮助。

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