Department of Natural Sciences, Bonn-Rhein-Sieg University of Applied Sciences, von-Liebig Str. 20, 53359 Rheinbach, Germany.
Institute for Functional Gene Analytics, Bonn-Rhein-Sieg University of Applied Sciences, Grantham Allee 20, 53757 Sankt Augustin, Germany.
Genes (Basel). 2024 Oct 5;15(10):1299. doi: 10.3390/genes15101299.
Modern forensic DNA quantitation assays provide information on the suitability of a DNA extract for a particular type of analysis, on the amount of sample to put into the analysis in order to yield an optimal (or best possible) result, and on the requirement for optional steps to improve the analysis. To achieve a high sensitivity and specificity, these assays are based on quantitative PCR (qPCR) and analyze target DNA loci that are present in multiple copies distributed across the genome. These target loci allow the determination of the amount of DNA, the degree of DNA degradation, and the proportion of DNA from male contributors. In addition, internal control DNA of a known amount is analyzed in order to inform about the presence of PCR inhibitors. These assays are nowadays provided as commercial kits that have been technically validated and are compatible with common qPCR instruments. In this review, the principles of forensic qPCR assays will be explained, followed by information on the nature of DNA loci targeted by modern forensic qPCR assays. Finally, we critically draw attention to the current trend of manufacturers not to disclose the exact nature of the target loci of their commercial kits.
现代法医 DNA 定量分析检测为以下方面提供信息:特定类型分析中 DNA 提取的适用性、为获得最佳(或尽可能好的)结果而应投入分析的样本量,以及可选步骤以改善分析的必要性。为了实现高灵敏度和特异性,这些检测基于定量聚合酶链式反应(qPCR),并分析分布在基因组中的多个拷贝的目标 DNA 基因座。这些目标基因座允许确定 DNA 量、DNA 降解程度以及来自男性供体的 DNA 比例。此外,还分析已知数量的内部对照 DNA,以了解是否存在 PCR 抑制剂。目前,这些检测以经过技术验证并与常见 qPCR 仪器兼容的商业试剂盒形式提供。在这篇综述中,将解释法医 qPCR 检测的原理,然后介绍现代法医 qPCR 检测中靶向的 DNA 基因座的性质。最后,我们批判性地提请注意制造商目前不披露其商业试剂盒的目标基因座的确切性质的趋势。