Nogami Hiroaki, Tsutsumi Hirofumi, Komuro Toshinobu, Mukoyama Rei
Department of Legal Medicine, Nihon University School of Dentistry, Chiyoda-ku, Tokyo, Japan.
Forensic Sci Int Genet. 2008 Sep;2(4):349-53. doi: 10.1016/j.fsigen.2008.05.001. Epub 2008 Jul 2.
Sex determination from dental pulp DNA was examined by loop-mediated isothermal amplification (LAMP) method. Amelogenin locus was analyzed for sex determination. A set of four specially designed primers was prepared based on database from Gene Bank, and loop primers were designed to shorten the analysis time. Analysis was performed using 32 dental pulp DNA samples removal from permanent teeth stored at room temperature for 1-25 years after extraction. The X allele was detected in approximately 32min with real-time turbidimeter and the Y allele was detected in approximately 34min. Analysis time was reduced to half when using loop primers. Visual detection was also possible as the amplified product showed white turbidity. Sex determination by LAMP method was rapid and simple, and it should prove useful in unknown bodies of mass disasters.
采用环介导等温扩增（LAMP）法对牙髓DNA进行性别鉴定。分析牙釉蛋白基因座以确定性别。根据基因库数据库制备了一组四个专门设计的引物，并设计了环引物以缩短分析时间。使用从室温下储存1 - 25年的恒牙中提取的32份牙髓DNA样本进行分析。使用实时浊度仪在约32分钟时检测到X等位基因，在约34分钟时检测到Y等位基因。使用环引物时，分析时间缩短了一半。由于扩增产物呈现白色浑浊，也可以进行目视检测。LAMP法进行性别鉴定快速且简单，在大规模灾害的无名尸体鉴定中应会证明有用。
