摘要
目的对2个钴胺素代谢障碍C型(Cobalamin C,cblC)家系进行基因变异分析,在明确变异的基础上建立针对MMACHC基因热点致病变异c.609G>A的聚合酶链反应-高分辨熔解曲线(high-resolution melting,HRM)快速筛查方法。方法提取先证者及父母外周血基因组DNA;采用PCR-Sanger测序技术对cblC家系的MMACHC基因进行变异分析;通过PCR-HRM技术筛查100名健康儿童MMACHC基因变异c.609G>A。结果经Sanger测序确认先证者1 MMACHC基因存在c.394C>T和c.609G>A复合杂合变异,先证者2 MMACHC基因存在c.482G>A和c.609G>A复合杂合变异。先证者及100名健康儿童PCR-HRM分析结果与Sanger测序结果一致。结论c.609G>A是MMACHC基因的热点致病变异,利用PCR-HRM技术对其进行筛查是cblC获得快速基因诊断有效方法。
Objective To carry out genetic testing for two families affected with cobalamin C(cblC)and establish a rapid method for the detection of a hotspot pathogenic variant c.609G>A of the MMACHC gene by using a high-resolution melting curve(PCR-HRM)method.Methods Genomic DNA was extracted from peripheral blood samples of the probands and their parents.Potential variants of the MMACHC gene was analyzed by Sanger sequencing.The c.609G>A variant of the MMACHC gene was screened among 100 healthy children with the PCR-HRM method.Results Sanger sequencing revealed that proband 1 carried compound heterozygous variants c.394C>T and c.609G>A of the MMACHC gene,while proband 2 carried compound heterozygous variants c.482G>A and c.609G>A of the same gene.PCR-HRM analysis of the two probands and the 100 healthy children were consistent with the Sanger sequencing.Conclusion c.609G>A is a hotspot pathogenic variant of the MMACHC gene.The diagnosis of cblC may be rapidly attained through detection by PCR-HRM.
作者
林书祥
王朝
张新杰
徐晓薇
邹倩倩
蔡春泉
张玉琴
舒剑波
Lin Shuxiang;Wang Chao;Zhang Xinjie;Xu Xiaowei;Zou Qianqian;Cai Chunquan;Zhang Yuqin;Shu Jianbo(Tianjin Pediatric Research Institute,Tianjin Children's Hospital,Tianjin 300134,China;Department of Neurology,Tianjin Children's Hospital,Tianjin 300134,China;Department of Neurosurgery,Tianjin Children's Hospital,Tianjin 300134,China)
出处
《中华医学遗传学杂志》
CAS
CSCD
2020年第7期759-763,共5页
Chinese Journal of Medical Genetics
基金
天津市自然科学基金(16JCQNJC11900)
国家自然科学基金(81771589)
天津市重大疾病防治科技重大专项(18ZXDBSY00170)。
