免疫磁珠法与差异裂解法检验混合样本的效果观察  

Comparative study of magnetic beads method and differential lysis method for the separation and detection of mixed cell samples

作　　者：李学博[1,2] 丁春丽[1] 宁淑华[3] 王清山[2] 封宇[2] 李红卫[2] 

机构地区：[1]山东政法学院山东省高校证据鉴识重点实验室,山东济南250014 [2]重庆市公安局物证鉴定中心,重庆400021 [3]重庆市公安局江北分局技术科,重庆400020

出　　处：《中国法医学杂志》2015年第3期249-251,255,共4页Chinese Journal of Forensic Medicine

基　　金：山东省自然科学基金资助项目(ZR2014HQ018);山东省高校证据鉴识重点实验室(山东政法学院)开放课题(2014-01);重庆市公安局科技攻关项目(2014-10)

摘　　要：目的分析应用免疫磁珠分离技术和差异裂解法对混合样本的DNA检验效果,评价其在法医学应用中的价值。方法制备含有不同数量精细胞与阴道上皮细胞混合悬液和植绒混合斑拭子,分别采用MOSPD3抗体、SPAG8抗体免疫磁珠法及差异裂解法对样本进行检验,观察分析不同比例的混合细胞悬液,以及不同保存时间植绒混合斑拭子DNA分型的正确率。结果当混合细胞悬液内精细胞与阴道上皮细胞比例约1∶10时,免疫磁珠法的正确率明显高于差异裂解法(P<0.01);当比例等于或高于1∶1时,两种方法正确率的差异无显著性意义(P>0.05)。植绒拭子混合斑的免疫磁珠法正确率明显低于差异裂解法(P<0.01);随混合斑保存时间的延长,差异裂解法正确率无明显下降(P>0.05)。结论免疫磁珠法适用于新鲜混合样本中精细胞的分离检验,且在精细胞含量较低时优势更明显,差异裂解法较适合陈旧混合斑的分离检验。Objective Results of antibody-magnetic beads method and differential lysis method for the separation of sperm in mixed cell samples were compared, to explore the potential feasibility in forensic practice. Methods Three different concentration mixed cell suspensions and simulated stains of sperm and vaginal epithelial cell was prepared. All the samples of mixed cell suspensions and simulated mixed stains were detected by antibody-magnetic beads method and differential lysis method respectively. The number of STR loci and the correct detection rate were calculated and compared. Results The correct detection rate of antibody-magnetic beads method was higher than differential lysis method when the number of sperm cells and vaginal epithelial cell was approximately 1 ： 10 （ P 〈 0. 01 ）. While there was no significant difference for the two methods when the number of sperm cells and vaginal epithelial cell was equal to or more than 1 ： 1 （P 〉 0.05 ）. The correct detection rate of antibody-magnetic beads method was lower than differential lysis method for the samples of simulated mixed stains in different storage time groups （P 〈 0. 01 ）. While there was no significant difference for the differential lysis method with the samples preservation time lasting （P 〉 0. 05 ）. Conclusion Antibody-magnetic beads method was effective for the fresh mixed samples especially when contains few sperm cells. Differential lysis method was more suitable for the old mixed samples.

关 键 词：法医物证学 免疫磁珠 混合斑 精子 DNA 

分 类 号：D919[医药卫生—法医学]