激光捕获显微分离技术分离多人混合精斑  

DNA profiling of spermatozoa mixture by low volume PCR combined with laser capture microdissection

作　　者：杨帆[1,2] 曾宪海[3] 丰蕾[1] 徐程[1,2] 赵禾苗[4] 李彩霞[1] 胡兰[1] 

机构地区：[1]公安部物证鉴定中心法医遗传学公安部重点实验室,北京100038 [2]中国人民公安大学,北京100038 [3]福建省公安厅,福建福州350003 [4]北京大学医学部,北京100191

出　　处：《中国法医学杂志》2014年第4期304-307,共4页Chinese Journal of Forensic Medicine

基　　金：国家自然科学基金资助项目(81202384);"十二五"国家科技支撑计划项目(2012BAK02B04)

摘　　要：目的利用激光捕获显微分离技术(LCM)分离多供体混合精斑。方法将3名自愿者供给的精液混合后,制备成混合精斑,并涂于覆膜玻片上。用PALM激光捕获显微分离系统切割并捕获样本中的单个精子,利用低体积扩增技术(LV-PCR)扩增并进行检测,并对分型结果进行统计。结果联用LCM和低体积扩增技术捕获样本中单个精子细胞进行扩增,可获得每位自愿者的Y-STR分型,分型成功率为73.15%。结论激光捕获显微分离联用低体积扩增技术可完成多供体混合精斑的分离检验。Objective To isolate and genotype the spermatozoa from multiple donors＇ semen mixture by laser capture microdissection （LCM） coupled to low volume PCR（LV-PCR）. Methods Semen samples from three donors were collected and mixed, then the cell pellet of the semen mixture stain was smeared onto PEN membrane slide. Under microscope, all single spermatozoa in sight were harvested by LCM. Finally, multiplex PCRs were performed using the on-chip LV-PCR and DNA profiles obtained were analyzed. Results DNA of each sperm cell captured by LCM was amplified by LV-PCR using AmpFISTR Y-filerTM Kits and DNA profiles of each contributors＇ from their mixture semen sample were successfully generated. The detection rate was up to 73.15%. Conclusion The combination of LCM and LV-PCR has been shown to be capable of discriminating sperm cells from multiple donors.

关 键 词：法医物证学 激光捕获显微分离技术 多人混合精斑 

分 类 号：D919[医药卫生—法医学]