高分辨率熔解曲线技术在筛查琥珀酸半醛脱氢酶缺陷症致病基因突变中的应用  

Application of high resolution melting curve in screening succinate semialdehyde dehydrogenase deficiency diseasecausing gene mutations

作　　者：董昭樱[1] 张新杰 支秀芳 范文轩[3] 张玉琴[3] 舒剑波[2] 蔡春泉[2] 李东[3] DONG Zhao-ying;ZHANG Xin-jie;ZHI Xiu-fang;FAN Wen-xuan;ZHANG Yu-qin;SHU Jian-bo;CAI Chun-quan;LI Dong(Department of Neurology,Tianjin Union Medical Centre,Tianjin 300121,Tianjin,China;Tianjin Pediatric Research Institute,Tianjin Key Laboratory of Birth Defects for Prevention and Treatment,Tianjin Children's Hospital·Children's Hospital of Tianjin University,Tianjin 300134,China;Department of Neurology,Tianjin Children's Hospital·Children's Hospital of Tianjin University,Tianjin 300134,China)

机构地区：[1]天津市人民医院神经内科,天津300121 [2]天津市儿童医院·天津大学儿童医院,天津市儿科研究所·天津市儿童出生缺陷防治重点实验室 [3]天津市儿童医院·天津大学儿童医院神经内科,天津300134

出　　处：《生物医学工程与临床》2022年第1期81-85,共5页Biomedical Engineering and Clinical Medicine

基　　金：国家自然科学基金资助项目(81771589);天津市重大疾病防治科技重大专项(18ZXDBSY00170);天津市卫生健康科技项目(ZC20120)。

摘　　要：目的对4例琥珀酸半醛脱氢酶缺陷症(SSADHD)患儿家系进行基因突变分析,在明确突变的基础上建立针对醛脱氢酶5家族成员A1基因(ALDH5A1)中常见致病基因突变位点c.1529C> T(p.S510F)的高分辨率熔解(HRM)曲线快速筛查方法。方法先证者4例SSADHD患儿年龄(5.3±2.2)个月,其中男性3例,女性1例。商品化试剂盒提取4例先证者及其亲属外周血基因组DNA;采用一代测序技术对患者家系的ALDH5A1进行基因突变分析;应用HRM曲线技术随机筛查80例患儿ALDH5A1上c.1529C> T位点突变。结果先证者均有致病基因突变c.1529C> T位点,其中2例为该位点纯合突变,2例杂合携带并伴有其他致病位点的杂合突变。先证者及其家属的HRM分析结果与测序结果一致。随机检测的80例患儿,年龄为(7.5±3.9)岁,其中男性48例,女性32例,均不携带该基因突变。结论推测c.1529C> T位点突变可能是津冀地区SSADHD患儿中ALDH5A1的常见致病基因突变位点,但其在人群中携带频率很低;利用HRM曲线技术可实现对该位点的快速筛查。Objective To analyze the gene mutations in 4 families of succinic semialdehyde dehydrogenase deficiency(SSADHD) children, and establish a high resolution melting(HRM) curve for rapid screening of common pathogenic mutation site c.1529 C > T(p.S510 F) in aldehyde dehydrogenase 5 family member A1 gene(ALDH5 A1). Methods The age of 4 SSADHD probands children was(5.3 ± 2.2) months, which included 3 males and 1 female. The commercial kits were used to extract genomic DNA from peripheral blood of probands and their relatives. First-generation sequencing technique was used to analyze the gene mutations of ALDH5 A1 in pedigree of the patients. HRM technology was used to randomly screen 80 children with c.1529 C > T mutations in ALDH5 A1. Results The probands showed pathogenic gene mutation c.1529 C > T, of which 2 were homozygous mutations in the locus, and 2 were heterozygous and carried heterozygous mutations in other pathogenic locus. The HRM analysis results of probands and their relatives were consistent with the sequencing results. None of the 80 randomly tested children[48 males and 32 females;aged(7.5 ± 3.9) years old] carried the gene mutation. Conclusion It is demonstrated that c. 1529 C > T could be a common pathogenic gene mutation of ALDH5 A1 in SSADHD patients in Tianjin and Hebei, but the carrying rate is very low in the population, and HRM curve can be used to achieve rapid screening of the locus.

关 键 词：高分辨率熔解曲线 琥珀酸半醛脱氢酶缺陷症 醛脱氢酶5家族成员A1基因(ALDH5A1) 儿童 

分 类 号：R589[医药卫生—内分泌] R725.9[医药卫生—内科学]