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PCR-SSO流式荧光微珠法HLA-A*02高分辨分型检测模棱两可结果的统计分析
引用本文:张容达,杜娟,马晓莉,张伯伟.PCR-SSO流式荧光微珠法HLA-A*02高分辨分型检测模棱两可结果的统计分析[J].中国组织工程研究与临床康复,2011,15(45).
作者姓名:张容达  杜娟  马晓莉  张伯伟
作者单位:1. 复旦大学物理系生物物理重点实验室,上海市,200433
2. 河南省红十字血液中心,河南省郑州市,450012
摘    要:背景:由于HLA的复杂多态性,实际分型过程中难免会出现一些结果判读模棱两可现象.目的:统计分析基于PCR-SSO 流式荧光微珠HLA-A*02高分辨分型方法的模棱两可组合结果种类、比例,探讨解决模棱两可解决方法,提高高分辨分型比例.方法:对河南骨髓库捐献者 1 100人份HLA分型结果进行统计分析,计数法进行分类统计分析软件给出的包含HLA-A*02:01\02:09模棱两可组合代码,计算百分比;与确认为A*02:09的基因多态性分布进行比对,找出与A*02:09关联性较强的等位基因,用补充实验进行复核、确认,建立A*02:01\02:09模棱两可组合的判读方法.结果与结论:1 039例有效数据中包含A*02:01\02:09模棱两可组合279例(279/1 039,26.853%),代码种类19种,检出比例较高的有DMDC,DYVK,GMDD,GMDE;34例确认为A*0209的基因多态性统计显示A*0209的出现与A*11:01 (0.147 1)B*07:02(0.264 7)、DRB1*01:01(0.264 7 )、DRB1*03:01(0.117 6)有较强关联,选出高风险标本38例,经检测外显子1~7,并用SBT复核确认有1例结果为A*02:09(代码组合为GMDC),其余241例未检出A*02:09.提示 A*02:09的出现与B*07:02、DRB1*01:01、DRB1*03:01有较强关联,重点对这些关联标本进行外显子1~7检测,可有效地检出A*02:09.通过统计分析制定合理的判定规则和必要的复核实验可有效地简化操作程序,节省时间,降低费用,提高高分辨比例.

关 键 词:HLA-A*0201/0209模棱两可  中华骨髓库  PCR-SSO  HLA分型  基因

Statistical analysis for ambiguous results of HLA-A*02 high-resolution genotyping by using PCR-SSO flow fluorescent bead method
Zhang Rong-da,Du Juan,Ma Xiao-li,Zhang Bo-wei.Statistical analysis for ambiguous results of HLA-A*02 high-resolution genotyping by using PCR-SSO flow fluorescent bead method[J].Journal of Clinical Rehabilitative Tissue Engineering Research,2011,15(45).
Authors:Zhang Rong-da  Du Juan  Ma Xiao-li  Zhang Bo-wei
Institution:Zhang Rong-da1,Du Juan2,Ma Xiao-li2,Zhang Bo-wei2 1Key Laboratory of Biophysics,Department of Physics,Fudan University,Shanghai 200433,China,2Henan Red-Cross Blood Center,Zhengzhou 450012,Henan Province
Abstract:BACKGROUND:Due to the complexity of the HLA typing system,ambiguous situations occur frequently during HLA typing of the donors.OBJECTIVE:To statistically analyze the ambiguous ratio and combined allele groups of HLA-A*02 high-resolution genotyping by using PCR-SSO flow fluorescent bead method,and to investigate the method to resolve the ambiguous results of HLA-A*02 high-resolution genotyping and improve the ratio of high-resolution genotyping.METHODS:1100 pieces of HLA genotyping were collected for statis...
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