Development and evaluation of an optimized Ancient DNA extraction method for femoral diaphyses and simulated heat-treated teeth.

IF 3.1
Guihong Liu, Shengqiu Qu, Qiushuo Wu, Yazi Zheng, Mengyu Tan, Jiaming Xue, Haoyan Jiang, Mengna Wu, Yuxuan Tan, Miao Liao, Jing Yan, Meili Lv, Lin Zhang, Weibo Liang
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Abstract

Extracting endogenous DNA from highly degraded hard tissue samples, such as bones and teeth, is crucial yet challenging for forensic human identification. Existing forensic methods and commercial kits for DNA extraction from bones and teeth often inefficient for old, poorly preserved, chemically treated, or heat-exposed samples. Ancient DNA (aDNA) extraction technology is highly effective in recovering degraded, short DNA fragments and has been successfully used to isolate DNA from skeletal remains and sediments dating back to tens of thousands of years. However, its direct application to individual identification and short tandem repeat (STR) analysis may require further adaptation to meet specific forensic requirements. Therefore, this study was conducted to optimize aDNA extraction techniques for developing a method tailored for highly degraded forensic samples. We developed a forensic aDNA-based extraction (FADE) method by refining the aDNA extraction protocol through optimization of lysis and purification conditions. The effectiveness of the developed method was validated using 8 forensic case samples (femoral diaphyses) and 20 simulated heat-treated teeth. For samples subjected to prolonged heat treatment, the FADE method significantly increased peak heights, with improvements of 30 % and 45 % in the 30- and 40-min heat-treatment groups, respectively, and yielded a greater number of amplified loci and alleles. The FADE method developed in this study is well-suited for forensic applications involving highly degraded femoral diaphyses and simulated heat-treated teeth, significantly enhancing the success rates of DNA analysis and providing an effective technical solution for complex forensic DNA extraction.

股骨骨干和模拟热处理牙齿古代DNA提取优化方法的开发与评价。
从高度降解的硬组织样本(如骨骼和牙齿)中提取内源性DNA对于法医人类鉴定至关重要,但也具有挑战性。从骨骼和牙齿中提取DNA的现有法医方法和商业试剂盒,对于陈旧的、保存不良的、经过化学处理的或暴露在高温下的样本来说,往往效率低下。古DNA (aDNA)提取技术在回收降解的短DNA片段方面非常有效,并已成功地用于从数万年前的骨骼遗骸和沉积物中分离DNA。然而,其直接应用于个体鉴定和短串联重复序列(STR)分析可能需要进一步适应,以满足特定的法医要求。因此,本研究旨在优化aDNA提取技术,以开发一种适合高度降解法医样本的方法。我们通过优化裂解和纯化条件,完善aDNA提取方案,开发了一种法医aDNA-based extraction (FADE)方法。采用8个法医病例样本(股骨干)和20个模拟热处理牙齿验证了该方法的有效性。对于经过长时间热处理的样品,FADE方法显著提高了峰高,在30分钟和40分钟热处理组分别提高了30 %和45 %,并且产生了更多的扩增位点和等位基因。本研究开发的FADE方法非常适合涉及高度降解的股骨干和模拟热处理牙齿的法医应用,显著提高了DNA分析的成功率,并为复杂的法医DNA提取提供了有效的技术解决方案。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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