目标序列捕获测序检出红细胞丙酮酸激酶缺乏症PKLR基因新的复合突变  

A PKLR Gene Novel Complex Mutation in Erythrocyte Pyruvate Kinase Deficiency Detected by Targeted Sequence Capture and Next Generation Sequencing

作　　者：李栋梁[1] 张静[2] 刘艳丽[1] 焦保全[1] 王志伟[1] 王友君[1] 李文静[1] 侯兰芬[1] 郭宏谋[1] 孙宇[1] 郭晓[1] 

机构地区：[1]解放军白求恩国际和平医院血液科,河北石家庄050082 [2]石家庄长城中西医结合医院内科,河北石家庄050000

出　　处：《中国实验血液学杂志》2015年第5期1464-1468,共5页Journal of Experimental Hematology

摘　　要：目的:探讨红细胞丙酮酸激酶缺乏症(pyruvate kinase deficiency,PKD)的分子发病机制。方法:应用目标序列捕获和高通量二代测序技术(next-generation sequencing,NGS)对临床拟诊PKD患儿的PKLR基因12个外显子及其侧翼序列进行测序,采用SIFT及PolyPhen-2数据库预测突变对蛋白质功能的影响,在确定患者致病基因型后,应用Sanger测序技术对此基因型进行验证。结果:NGS结果显示,患儿PKLR基因存在罕见的双重杂合突变:第5外显子661G>A(Asp221Asn)和第10外显子1528C>T(Arg510Ter),导致该基因的第221位氨基酸由天冬氨酸突变为天冬酰胺,第510位氨基酸由精氨酸突变为终止密码子,使PKLR基因氨基酸链编码提前终止;Sanger测序技术进一步验证了该双重突变的存在。检索相关文献及数据库显示,这两种突变在人群中的发生率极低,蛋白质功能预测均显示为有害。结论:PKLR基因661 G>A与1528 C>T双重杂合突变是该PKD患儿的分子发病机制,PKD患者同时存在上述复合突变的报道在国内外尚属首次。Objective: To explore the molecular mechanism of erythrocyte pyruvate kinase deficiency( PKD).Methods: Targeted sequence capture and next-generation sequencing( NGS) were used to detect the regions of exon and exon-intron boundarie of PKLR gene in a clinical suspected PKD patient. The protein function of mutant gene was forecasted by the SIFT and PolyPhen-2 datebank,after the mutation of PKLR gene in the patient was detected by the NGS technology,its genotype was confirmed by Sanger sequencing. Results: The patient was found to have peculiar double heterozygous mutations: 661 G > A( Asp221Asn) of exon 5 and 1528 C > T( Arg510Ter) of exon 10,resulting in amino acid substitution Asp221 Asn and Arg510 Ter,these mutations were also further confirmed by Sanger sequencing.The complex mutations were infrequent and each of them was able to cause diseases. Conclusion: The complex mutations of both 661 G > A and 1528 C > T of PKLR gene are the molecular mechanism of PKD. Simultaneous existance of above-mentioned complex mutations in PDK patient was never been previously reported at home and abroad.

关 键 词：基因 突变 丙酮酸激酶 目标序列捕获 二代测序 

分 类 号：R725.5[医药卫生—儿科]