微量口腔脱落细胞检材的DNA检验

目的寻求提高微量口腔脱落细胞检材的DNA检验成功率的简便有效的提取方法。方法对不同载体上的100份微量口腔脱落细胞检材采用小体积Chelex-100法提取DNA,在ABI7500型荧光定量PCR仪上进行定量,同时用IdentifilerTM复合扩增系统扩增,在ABI3130遗传分析仪上进行STR分型。结果从25根饮料吸管上提取的DNA量在0.72～116.7.8ng,16个水杯杯缘提取的DNA量在2.15-142.5ng,31个饮料瓶(罐)口提取的DNA量在1～34.65ng,10根筷子上提取的DNA量在3.35～26.6ng,12个果核中提取的DNA量在0.294～21.4ng,6份吃剩的骨头中提取的DNA量在0.88～5.88ng。100份检材性别及9个以上STR位点分型成功率平均为59.38%。除了使用者的个人原因外,检材的提取送检方式、检材的质地、饮料的性质对提取的DNA量有显著影响,是否加蛋白酶K对提取的DNA量无显著影响。结论采用小体积Chelex-100法可对60%左右的微量口腔脱落细胞检材提取DNA进行STR分型。

DNA typing of minimal oral epithelial cell samples

Objective To seek simple and cost-effective extraction technique to improve multiplex STR amplification from minimal oral epithelial cell samples.Methods One hundred DNA samples of oral epithelial cells extracted by mini system Chelex-100 method were quantitated by ABI 7500 Real Time System and were then typed with IdentifilerTM system in ABI 3130 Genetic Analyzer.Results The DNA contents of different categories of samples were as followings:suck pipes(0.72-116.78 ng),cup edges(2.15-142.5 ng),mouths of drink bottle(1-34.65 ng),chopsticks(3.35-26.6 ng),fruit cores(0.294-21.4 ng),and poultry bones(0.88-5.88 ng).The mean successful typing rate for gender and more than 9 STR loci was about 59.38%.Except the addition or no addition of proteinase K to the samples,all other factors-C users' variation,sample extraction methods,and qualities and properties of the samples had considerable effects on the contents of extracted DNA.Conclusion Successful STR typing can be achieved in about 60% minimal oral epithelial cell DNA samples extracted by mini Chelex system.