口腔拭子DNA检验的实时定量研究  

Real time PCR quantificational study of DNA extracted from oral swabs

作　　者：杨电[1] 李越[1] 刘超[1] 陈玲[2] 

机构地区：[1]广东广州市刑事科学技术研究所,510030 [2]南方医科大学法医教研室,广东广州510515

出　　处：《刑事技术》2007年第4期23-25,共3页Forensic Science and Technology

摘　　要：目的寻求提高口腔拭子的DNA检验效率的方法。方法对来源于不同个体或同一个体不同擦拭次数的口腔拭子用Chelex-100或磁珠法提取的DNA用实时定量PCR技术进行定量,同时用Identifiler复合扩增系统在ABI3100遗传分析仪上对这些DNA样品进行STR分型。结果在建立的Identifiler系统8μl扩增体系中,3个月内的口腔拭子用Chelex-100法提取的DNA模板1μl量较3μl量扩增效果好,磁珠法提取的DNA模板用量大小对复合扩增检测影响较小。结论用棉签擦拭颊粘膜5次制备口腔拭子,取其头部的约1/4用200μlChelex-100法提取DNA,然后用1μl模板进行复合扩增,是提高口腔拭子的DNA检验效率的简便可行的方法,但陈旧口腔拭子用磁珠法提取更能保证复合扩增分型成功。Objective To study how to improve multiplex STR amplification efficiency from oral swabs DNA. Method Different oral swab DNA extracted by Chelex-100 method or DNA IQ system magnetic bead method were quantitated by real time PCR technique, and typed with Identifiler systems in 3100 Genetic Analyzer. Results In the 8μl Identifiler amplification system established, the amplification of 1μl template was more efficient than 3μl for the DNA extracted from oral swabs within 3 months by Chelex-100 method. However, the quantity of DNA template extracted by DNA IQ system exerted little effects on the amplification efficiency. Conclusion It is simple and operative method to improve the DNA analysis efficiency extracted from oral swabs by swabbing oral mucous membrane 5 times firstly, then extracting DNA from 1/4 vertical swab by 200μl Chelex-100 method, and amplifying with 1μ template finally. The DNA of stale oral swabs extracted by DNA IQ system could be amplified more effectively.

关 键 词：口腔拭子 实时定量PCR技术 DNA定量 STR复合扩增 

分 类 号：D919.2[医药卫生—法医学]