【摘要】 目的 建立体细胞GPA突变频率的检测方法，研究环境基因毒素如电离辐射等对人体的损伤。方法 用自制3株对M和N型血型糖蛋白A(Glycophorin A,GPA)的特异单克隆抗体：3G4(抗M)，6A8(抗N)和3C5(抗MN)，进行荧光素和生物素标记，与甲醛和/或二甲亚胺固定的人外周血红细胞结合，在流式细胞上以荧光强度分选出正常和变异红细胞，分析MN型个体红细胞突变性质(即MN→MO、MM、NO、NN)，计算突变频率。结果 建立了GPA分析法1Wa、1Wb和2Wa法，1Wa、1Wb法可分别检测MN血型个体红细胞的MO、MM突变细胞群和NO、NN突变细胞群，2Wa法可将MN型个体红细胞分选出MO、M M、NO、NN型4种突变细胞，并确定变异细胞频率。结论 建立的方法可以用于人体细胞突变的检测。

Development of a humansomatic mutation detection

    method--GP A assay

MAO Jianping, DONG Yan,LIU Bin, etal.Biochemistry Laboratory,Beij ing Institute of Radiation Medicine,Beijing 100850, China

【Abstract】 Objective To study thedamage to human body caused by environmental radiation,and supervise the somaticmutations. Methods Three monoclonal antibodies specific to M-type(3G4),N-ty pe(6A8), andMN-type (3C5) of glycophorin A,respectively,were prepared.Fluores c ence or biotinconjugated antibodies were bound specifically to formalin and/or dimethyl suberimidatefixed erythrocytes.M,MN,and N type cells were divided by c ytometry to demonstrate theerythrocyte mutation characteristics (MN→MO,MM,NO,N N) and give out the variantfrequency. Results 1Wa,1Wb and 2Wa meth ods of GPA assay were developed.Erythrocytes of MNtype individuals could be sep a rated to normal and single locus variant groups by 1Wmethods and they could be sor ted as normal (MN),single gene deletion mutants (MO,NO),homozygous mutants (MM,N N) cell groups by 2Wa method. Conclusion The assay isapplicable to evaluationg the frequency of variant erythrocytes from human somaticmutation.

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