人骨骼及牙齿DNA提取方法的比较和优化

目的人骨骼和牙齿DNA提取方法的比较和优化。方法收集18份不同个体的长骨、30颗磨牙和同一个体2根股骨、8颗磨牙。利用TissueLyser-Ⅱ组织破碎仪和PreFiler Express BTA^TM法医DNA提取试剂盒（BTA法）,应用Automate Express^TM自动化法医DNA提取系统提取DNA,进行STR分型,与脱钙法进行比较,并进行实验条件优化。结果用TissueLyser-Ⅱ结合BTA法,约2.5h即可完成骨骼和牙齿的DNA提取,分型成功率分别为94.4%和96.7%。与脱钙法比较,两种方法获得DNA质量浓度和检出率比较接近（P〈0.05）,但BTA法在操作过程方面更具优势。最佳样本量为100mg,消化时间为2h。结论采用TissueLyser-Ⅱ组织破碎仪结合BTA法对骨骼和牙齿进行DNA提取和分型检验,能满足实际检案的要求,可在法医学实践中选择使用。

The comparison and improvement of forensic DNA extraction method for bones and teeth

Objective To compare and improve forensic DNA extraction method from bones and teeth samples. Methods Samples 18 bones and 30 teeth were acquired from skeletonized remains and triturated by TissueLyser-II. DNA materials were extracted from the triturated samples with PreFiler Express BTATM forensic DNA extraction kit by Automate ExpressTM forensic DNA extraction system （BTA method） and decalcification method. DNA was quantified and followed by genotyping. The BTA method optimal samples size and paralysis time were determined by two femurs and eight molars from the same individual. Results DNA extraction from bones and teeth were completed in 2. 5 hours using TissueLyser-II and the BTA method. The quality of extracted DNA concentration is high for 18 bones and 30 teeth in the test, the same sample at different methods there were no significant difference between the same weight （ P 〈 0. 05 ）, but the BTA method has more advantages in operation process. The success rate of bones and teeth STR typing were 94.4% and 96. 7% using the BTA method, respectively. The results showed the optimal samples size 100mg and paralysis time 2h. Conclusion The BTA method is an efficient method to extract DNA from bones and teeth, which are simple, can significantly improve the success rate of bones and teeth DNA test and can be applied in forensic practice.