葡萄糖⁃6⁃磷酸脱氢酶基因突变永生化淋巴细胞系的建立  

Establishment of immortalized lymphocyte cell lines with mutation of the glucose⁃6⁃phosphate dehydrogenase gene

作　　者：贾峥[1] 张艳艳 孙楠[1] 张文新 高飞[1] 孙晶 黄杰[1] 曲守方[1] JIA Zheng;ZHANG Yanyan;SUN Nan;ZHANG Wenxin;GAO Fei;SUN Jing;HUANG Jie;QU Shoufang(National Institutes for Food and Drug Control,Beijing,China,100050;MGI Tech Co.,Ltd,Shenzhen,China,518083)

机构地区：[1]中国食品药品检定研究院体外诊断试剂检定所,北京100050 [2]深圳华大智造科技有限公司,深圳518083

出　　处：《分子诊断与治疗杂志》2020年第4期428-432,共5页Journal of Molecular Diagnostics and Therapy

基　　金：生殖健康及重大出生缺陷防控研究(2016YFC1000300)。

摘　　要：目的收集27例葡萄糖⁃6⁃磷酸脱氢酶基因突变的临床外周血样本,建立永生化淋巴细胞系。方法采用葡萄糖⁃6⁃磷酸脱氢酶基因突变检测试剂盒(荧光PCR熔解曲线法)筛查临床样本的突变类型,然后采集外周血,进行永生化淋巴细胞系的转化。提取建系成功的淋巴细胞系的基因组DNA,并用建立的二代测序法进行验证。结果成功建立了27例葡萄糖⁃6⁃磷酸脱氢酶永生化淋巴细胞系样本,包括14种不同纯合和杂合突变类型,验证结果表明永生化淋巴细胞系的基因突变类型与临床样本原来标示的突变类型一致。结论本研究建立的葡萄糖⁃6⁃磷酸脱氢酶基因突变永生化淋巴细胞系具有稳定的遗传信息,可以作为国家参考品的资源,用于评价葡萄糖⁃6⁃磷酸脱氢酶基因突变试剂盒的性能。Objective To establish immortalized lymphocyte cell lines by collecting 27 clinical peripheral blood samples harboring glucose⁃6⁃phosphate dehydrogenase(G6PD)gene mutations.Methods The glucose⁃6⁃phosphate dehydrogenase gene mutation detection kit(fluorescence PCR melting curve method)was used to screen the mutation type of clinical samples,and then peripheral blood was collected for transformation of immortalized lymphocyte lines.The genomic DNA of the successfully established lymphocyte line was extracted and verified by the next⁃generation sequencing method.Results We established 27 immortalized lymphocyte cell lines,including 14 different homozygous or heterozygous G6PD mutation types.The genetic mutation types of immortalized lymphocyte lines were the same as those originally indicated in clinical samples.Conclusion The established immortalized lymphocyte cell lines with G6PD mutations in this study have stable genetic information and can be used as a national reference resource to evaluate the performance of the G6PD gene mutation kit.

关 键 词：葡萄糖⁃6⁃磷酸脱氢酶 基因突变 永生化细胞系 

分 类 号：R394[医药卫生—医学遗传学]