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重组酶聚合酶放大法医短串重复位置的重组酶

Sonja Škevin¹, Liesl De Keyzer¹, Lynn De Waele¹

¹Laboratory of Pharmaceutical Biotechnology, Ghent University, Ottergemsesteenweg 460, Gent, 9000, Belgium.

Scientific reports

|November 18, 2025

概括

重组酶聚合酶放大 (RPA) 显示出对快速,便携式法医DNA分析的承诺. 虽然单复数放大产生了准确的短串重复 (STR) 配置文件,但多复数放大仍然是一个挑战.

科学领域:

法医科学法医科学法医科学
分子生物学分子生物学
生物技术是生物技术.

背景情况:

短串联重复 (STR) 对于法医DNA鉴定至关重要.
目前的方法,如毛细电泳 (CE) 和下一代测序 (NGS) 是集中和耗时的.
需要快速,便携且具有成本效益的STR基因型鉴定.

研究的目的:

评估复合酶聚合酶放大 (RPA) 用于放大法医相关的STR位点.
评估RPA适用于小型和便携式STR基因型定型设备的适用性.
为了将RPA性能与CE和测序等既定方法进行比较.

主要方法:

使用RPA在单复式和多复式格式中对13个CODIS STR位点进行放大.
通过CE,Illumina测序和牛津纳米孔技术 (ONT) 测序对RPA安普利康的分析.
评估测定灵敏度到62ppg的DNA输入.

主要成果:

Singleplex RPA成功地放大了STR位点,通过CE,Illumina和ONT获得了完整和正确的配置文件.
准确的基因型定型在大多数位置只需62 pg的DNA输入就能实现.
多重RPA放大导致不完整或不正确的STR配置文件,表明同时放大具有挑战性.

结论:

在法医应用中,RPA是一种可行的异热放大方法,用于单复 STR 基因定型.
RPA为开发快速和便携式DNA识别工具提供了潜力.
需要进一步优化,以克服多重RPA的挑战,以实现全面的法医DNA分析.

关键词:

毛细管电泳可以通过毛细管电泳进行.法医遗传学法医遗传学大规模并行测序.微卫星是微观卫星中的一个.纳米孔测序的测序复合酶聚合酶放大复合酶的放大.短串联重复的重复.

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