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纸质出版:2012
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李艮平,李彩霞,郝建文,万立华,胡兰,韩俊平,杨帆.激光捕获显微切割技术诊断石蜡切片中早期胚胎成分与亲权关系的研究[J].陆军军医大学学报(原第三军医大学学报),2012,34(15):1501-1504.
Li Genping,Li Caixia,Hao Jianwen,Wan Lihua,Hu Lan,Han Junping,Yang Fan.Separation of chorionic villi from paraffin-embedded early abortion materials and confirmation of parent-child relationship by laser capture microdissection[J].J Amry Med Univ (J Third Mil Med Univ),2012,34(15):1501-1504.
李艮平,李彩霞,郝建文,万立华,胡兰,韩俊平,杨帆.激光捕获显微切割技术诊断石蜡切片中早期胚胎成分与亲权关系的研究[J].陆军军医大学学报(原第三军医大学学报),2012,34(15):1501-1504. DOI:
Li Genping,Li Caixia,Hao Jianwen,Wan Lihua,Hu Lan,Han Junping,Yang Fan.Separation of chorionic villi from paraffin-embedded early abortion materials and confirmation of parent-child relationship by laser capture microdissection[J].J Amry Med Univ (J Third Mil Med Univ),2012,34(15):1501-1504. DOI:
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。 方法 通过激光捕获显微技术捕获石蜡切片上胚胎绒毛细胞,设置20、30、40个细胞数目组,分别捕获不同数目绒毛细胞,利用低体积扩增仪进行1.5 μl体系PCR扩增,采用3130遗传分析仪得出胚胎个体分型图谱,并比较各细胞数目组胚胎个体识别的检出率、等位基因丢失率和非特异性扩增情况。最后进行实际案例应用。 结果 捕获20个胚胎绒毛细胞即可获得胚胎个体分型,40个细胞数目组的胚胎分型检出率最高,等位基因丢失率和非特异性扩增率最低。20个细胞数目反之。 结论 激光捕获显微切割技术结合低体积扩增技术可应用于石蜡切片,实现胚胎与母体成分的精确分离并确证亲权关系。
Objective It remains difficult to genotype the mixture of maternal and embryo from early abortion tissues to investigate forensic evidence and confirm parent-child relationship. Our study was aimed to confirm the parent-child relationship by using laser capture microdissection (LCM) to capture the embryonic villi cells in paraffin-embedded section for genotyping embryonic individual via ultralow volume PCR amplification. Methods Embryonic villi cells in the paraffin sections was captured by LCM technology. With the setting of cell numbers at 20
30 and 40 cells as a 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 also applied on an actual case. Results Embryonic villi cells in the paraffin sections were captured by LCM technology. With the setting of cell numbers at 20
30 and 40 cells for group
villus cells of different cell numbers were captured and subjected to PCR amplification at low volume of 1.5 μl
and embryonic genotyping maps were obtained. The rate of embryo genotyping in the group of 40 cells was the highest whereas the rate of allele loss and non-specific amplification was the lowest. Vise versa for the group of 20 cells. Conclusion LCM combined with the low volume amplification technique can be applied to paraffin-embedded tissue sections to achieve accurate separation of the embryo and maternal components and obtain conclusive evidence for parental rights.
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