激光捕获显微切割技术诊断石蜡切片中早期胚胎成份与亲权关系的研究.DOC

激光捕获显微切割技术石蜡切片中胚胎成基础医学院法医学教研室1； 100038 北京,公安部物证鉴定中心2，法医遗传学公安部重点实验室北京3, 400014 重庆，重庆市公安局4；100038 北京,中国人民公安大学5） [摘要] 目的 通过激光捕获显微切割技术捕获石蜡切片中胚胎绒毛细胞,进行低体积扩增，实现胚胎个体分型,证实亲子关系。R89 ［文献标识码］ A Separation of chorionic villi from paraffin-embedded tissue section of early abortion materials by laser capture microdissection Li Genping1, LI Caixia2,3, Hao Jianwen 4，Wan Lihua1，HU Lan2,3, Han Junping5，Yang Fan5 (1hongqing Medical University，Chongqing400016;2Institute of Forensic Science of Ministry of Public Security; 3Key Laboratory of Forensic Genetics, Ministry of Public Security, Beijing,100038; 4ChongQing public security bureau，Chongqing400014;5Chinese People’s Public University, Beijing 100038, China) ［Abstract］ Objective It remains difficult to genotype the mixture of maternal and embryo from early abortion tissues for investigating forensic evidence and confirming parent-child relationship. The current study was designed to confirm the parent-child relationship by using LCM to capture the embryonic villi cells in paraffin section for geneotyping embryonic individual via outlow volume PCR amplification. Methods Embryonic villi cells in the paraffin sections was captured by laser capture microdissection technology. With the setting of cell numbers at 20, 30 and 40 per cell group, villus cells of different cell numbers were captured and subjected to PCR amplification at low volume of 1.5 μL by using 3130 Genetic Analysis instrument to yield embryonic genotyping maps. The artificial alleles, detection rate and allelic dropout rate of each group were compared. The technology was applied on an actual case. Results Embryonic villi cells in the paraffin sections was captured by laser capture microdissection technology. With the setting of cell numbers at 20, 30 and 40 per cell group, villus cells of different cell numbers were captured and subjected to PCR amplification at low volume of 1.5 μl by using 3130 Genetic Analysis instrument to yield embryonic genotyping