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采用 Magelia 平台上的 GlobalFiler™ IQC PCR 扩增试剂盒进行低投入法医 DNA 样本分析的创新方法。

An innovative approach for low input forensic DNA sample analysis using the GlobalFiler™ IQC PCR amplification Kit on the Magelia® platform.

机构信息

Institut de recherche criminelle de la gendarmerie nationale (IRCGN), Cergy-Pontoise, 95000, France; Centre national de recherche en génomique humaine (CNRGH), Commissariat à l'énergie atomique et aux énergies alternatives (CEA), Evry-Courcouronnes 91000, France.

Inorevia, Bagneux 92220, France.

出版信息

Forensic Sci Int Genet. 2024 Sep;72:103093. doi: 10.1016/j.fsigen.2024.103093. Epub 2024 Jul 7.

Abstract

Short Tandem Repeat (STR) markers have been the gold standard for human identification testing in the forensic field for the last few decades. The GlobalFiler™ IQC PCR amplification Kit has shown sensitivity, high power of discrimination and is therefore widely used. Samples with limited DNA quantities remain a significant hurdle for streamlined human forensic identification. Reaction volume reduction in a closed system paired with automation can provide solutions to secure DNA profiles when routine methods fall short. We automated and optimized the GlobalFiler IQC PCR Amplification Kit on the Magelia®, a closed molecular biology platform, to test whether reaction volume reduction in a confined automated system would improve signal and sensitivity. We evaluated the platform's performance using reference and real casework samples (blood, cigarette butt, saliva and touch DNA) in the context of a 5-fold volume reduction when compared to the routine protocol. This strategy showed distinct advantages over standard treatment, notably increased signal for lower DNA inputs. Importantly, negative casework samples through routine treatment yielded "usable" DNA profiles after amplification using this strategy. This novel approach represents a first proof of concept for a method enabling users to treat limited samples, or to partition routine samples for multiple analyses.

摘要

短串联重复序列(STR)标记物在过去几十年一直是法医领域人类身份识别检测的金标准。GlobalFiler™ IQC PCR 扩增试剂盒具有灵敏度高、高分辨力的特点,因此被广泛应用。对于有限量 DNA 的样本,仍然是简化人类法医鉴定的一个重大障碍。在常规方法失败的情况下,减少封闭系统中的反应体积并结合自动化可以提供获取 DNA 图谱的解决方案。我们在 Magelia®上自动化并优化了 GlobalFiler IQC PCR 扩增试剂盒,以测试在封闭自动化系统中减少反应体积是否会提高信号和灵敏度。我们使用参考样本和实际案例样本(血液、烟头、唾液和接触性 DNA)来评估该平台的性能,与常规方案相比,反应体积减少了 5 倍。与标准处理相比,该策略具有明显的优势,尤其是在较低的 DNA 输入时增加了信号。重要的是,通过常规处理的阴性案例样本在使用该策略进行扩增后,产生了“可用”的 DNA 图谱。这种新方法代表了一种方法的初步概念验证,该方法可以让用户处理有限的样本,或者将常规样本分割用于多个分析。

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