DIAGNOSTIC METHODS BY DNA ANALYSIS OF DRIED BLOOD SPOTS AND CLASSIFICATION OF INHERITED DISEASE BY MUTANT GENOTYPES.

通过干血斑 DNA 分析的诊断方法和通过突变基因型对遗传性疾病进行分类。

• 批准号: 02557041
• 负责人: NARISAWA K
• 金额: $ 11.07万
• 依托单位: TOHOKU UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Developmental Scientific Research (B)
• 财政年份: 1990
• 资助国家: 日本
• 起止时间: 1990 至 1992
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-02557041/
• 关键词: MOLECULAR GENETICS; GENE CLONING; GENE ANALYSIS; GENETIC DIAGNOSIS; INHERITED DISEASE; 遺伝子診断; 遺伝子変異; 中鎖アシルCoA脱水素酵素; メチルマロン酸血症

1. Medium chain acyl-CoA dehydrogenase(MCAD) deficiency is an autosomal recessive disorder which is known to cause Reye-like syndrome in children and sudden infant death. A point mutation of lysine(329)-to-glutamic acid(329)substitution in the MCAD gene was identified as the most common mutation in patients with MCAD deficiency. This mutation is responsible for about 90% of mutant MCAD alleles in Caucasians and now be detected by DNA diagnostic methods using Guthrie cards. Screening for the mutation among newborns in England, Australia, and United States of America indicates the prevalence of carriers to be 1 of 40-107, suggesting the high incidence of the mutation.2. Nonketotic hyperglycinemia(NKH) is an autosomal recessive metabolic disorder. We have cloned and characterized a human glycine decarboxylase cDNA responsible for NKH. This clone was 3,705 bp in length and encoded 1,020 amino acids. NKH is a disorder with high incidence in northern Finland. To understand the genetic backgr … More ound of this high incidence, we examined the glycine decarboxylase in a typical case of NKH. Structural analysis of glycine decarboxylase mRNA from the patient revealed a single nucleotide substitution from G to T in the protein coding region, which resulted in an amino acid alteration from Ser(564) to Ile(564). The patient was homozygous for this mutation. Furthermore, this mutation was present in 70% of glycine decarboxylase gene alleles in Finnish patients with NKH, whereas it was not found in 20 alleles of non-Finnish patients. The results suggest that this mutation is responsible for the high incidence of NKH in Finland.3. Phenylketonuria(PKU) is an autosomal recessive disorder caused by a deficiency of the hepatic phenylalanine hydroxylase. We have developed a simple rapid DNA screening test that allows us to analyze seven known PKU mutations in Orientals, This method is based on an allele specific PCR of the genomic DNA extracted from Guthrie cards and provides an easy method of simultaneously analyzing plural mutations. Less

1.中链酰基辅酶A脱氢酶（MCAD）缺乏症是一种常染色体隐性遗传疾病，已知可引起儿童Reye样综合征和婴儿猝死。MCAD基因中赖氨酸（329）-谷氨酸（329）取代的点突变被确定为MCAD缺乏症患者中最常见的突变。这种突变导致高加索人中约90%的突变MCAD等位基因，现在可以通过使用古特里卡的DNA诊断方法检测到。在英国、澳大利亚和美国的新生儿中进行的突变筛查表明，携带者的患病率为40-107分之一，这表明突变的发生率很高。非酮性高甘氨酸血症（NKH）是一种常染色体隐性遗传代谢紊乱。我们已经克隆并鉴定了负责NKH的人甘氨酸脱羧酶cDNA。该克隆全长3,705 bp，编码1,020个氨基酸。NKH是一种在芬兰北方发病率很高的疾病。为了了解遗传背景， ...更多信息 鉴于这种高发病率，我们检测了一例典型NKH的甘氨酸脱羧酶。甘氨酸脱羧酶mRNA的结构分析显示，在蛋白质编码区的一个单核苷酸从G到T的取代，这导致氨基酸从Ser（564）到Ile（564）的改变。该患者为该突变的纯合子。此外，这种突变存在于70%的甘氨酸脱羧酶基因等位基因在芬兰NKH患者，而它没有被发现在20个等位基因的非芬兰患者。结果表明，这种突变是负责高发病率的NKH在芬兰。苯丙酮尿症（PKU）是一种常染色体隐性遗传病，由肝脏苯丙氨酸羟化酶缺乏引起。我们建立了一种简单快速的DNA筛查方法，可以同时分析东方人的7个已知PKU突变。该方法基于从古特里卡中提取的基因组DNA的等位基因特异性PCR，提供了一种简单的同时分析多个突变的方法。少

项目成果

KURE,S,NARISAWA,K.TADA,K.: "ENZYMATIC DIAGNOSIS OF NONKETOTIC HYPERGLYCINEMIA;A NOVEL ASSAY OF GLYCINE CLEACAGE SYSTEM ACTIVITY USING LYMPHOBLASTS TRANSFORMED BY EPSTEINーBARR VIRUS." J.PEDIATR.

KURE,S,NARISAWA,K.TADA,K.：“非酮症高甘氨酸血症的酶促诊断；利用 EPSTEIN-BARR 病毒转化的淋巴母细胞对甘氨酸裂解系统活性进行的新颖测定。”

KURE,S.: "STRUCTUAL AND EXPRESSION ANALYSIS OF NORMAL AND MUTANT mRNA ENCODING GLYCINE DECARBOXYLASE:THREE BASE DELETION IN IN mRNA CAUSES NONKETOTIC HYPERGLYCINEMIA." BIOCHEM.BIOPHYS.RES.COMMUN.

KURE,S.：“编码甘氨酸脱羧酶的正常和突变 mRNA 的结构和表达分析：mRNA 中的三碱基缺失导致非酮性高甘氨酸血症。”

Kure,S.et.al.: "Identification of a common mutation in Finnish patients with nonketotic hyperglycinemia." J.Clin.Invest.90. 160-164 (1992)

Kure,S.et.al.：“芬兰非酮症高甘氨酸血症患者常见突变的鉴定。”

Matsubara,Y.: "Prevalence of K329E mutation in medium-chain acyl-CoA dehydrogenase gene determined from Guthrie cards." LANCET. 338. 552-553 (1991)

Matsubara,Y.：“根据 Guthrie 卡确定中链酰基辅酶 A 脱氢酶基因中 K329E 突变的发生率。”

MATSUBARA,Y.et.al.: "Prevalence of K329E mutation in medium-chain acyl-CoA dehydrogenase gene determined from Guthrie Cards." LANCET. 338. 552-553 (1991)

MATSUBARA,Y.et.al.：“根据 Guthrie Cards 确定的中链酰基辅酶 A 脱氢酶基因中 K329E 突变的发生率。”