不同血型个体混合细胞的免疫标识及DNA检验  

Separation of Mixed Cell Samples of Different Blood Groups by Immuno-labelling and DNA Typing

作　　者：王清山[1,2] 李学博[1] 李红卫[1,2] 

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

出　　处：《中国司法鉴定》2015年第5期44-47,共4页Chinese Journal of Forensic Sciences

基　　金：山东省自然科学基金资助项目(ZR2014HQ018);上海市法医学重点实验室开放课题(KF1510);山东省高校证据鉴识重点实验室(山东政法学院)开放课题(KFKT-201401);重庆市公安局科技攻关项目(G2014-10)

摘　　要：目的应用免疫技术标识不同ABO血型个体的上皮细胞,建立混合物证分离检验鉴定方法。方法取正常人体口腔上皮细胞,制备不同血型个体的混合口腔上皮细胞,经免疫组化染色,根据细胞膜的呈色差异,利用激光捕获显微切割目的细胞5个、10个、20个,所获样本进行多位点低体积复合扩增,扩增产物用ABI3130XL型DNA测序仪检测,Gene Mapper ID software vs.3.2软件分析,以获得13个以上STR分型为有效分型(各等位基因峰高大于50RFU)。结果根据免疫组化结果,可对ABO血型个体的混合细胞中的某单一血型细胞进行分离检验。DNA有效分型检出率随着切割回收细胞数的增加而提高,切割20个细胞时样本的DNA检出率达到90%以上。结论利用免疫标识技术可以分离不同ABO血型个体的上皮细胞,对混合样本的分离检验有重要价值。Objective To establish a new method for separating mixed epithelial cell mixtures from individuals of different ABO blood groups by immunological method. Method Mixed oral epithelial cell samples from individuals of different ABO blood groups were prepared. The cells were distinguished by immunohistochemical staining method. Three groups of samples which contained 5, 10 and 20 oral epithelial cells were captured respectively by laser capture micro-dissection. All the samples were then amplified by low volume amplification techniques. The PCR products were separated and analyzed with a 3130XL genetic analyzer and GeneMapper ID software version 3.2. The number of correctly amplified gene loci was calculated. Results The cells of a single blood group can be separated from the mixture by the established method. The detection rate of DNA typing increased with the number of oral epithelial cells. When 20 oral epithelial ceils were captured and amplified, the rate of detection reached 90%. Conclusion Immunohistochemical staining method is effective for separating the mixed samples from individuals of different ABO blood groups and can be utilized in forensic practice.

关 键 词：法医遗传学 混合细胞 DNA STR 

分 类 号：DF795.4[医药卫生—法医学]