Rapid Detection of Known Mutations and Its Application to Carrie Testing

已知突变的快速检测及其在Carrie检测中的应用

• 批准号: 06557046
• 负责人: NARISAWA Kuniaki
• 金额: $ 8.06万
• 依托单位: Tohoku University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (A)
• 财政年份: 1994
• 资助国家: 日本
• 起止时间: 1994 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-06557046/
• 关键词: Cloning; Mutation; Allele Specific PCR; Multiplex ASPCR; Holocarboxylase Synthetase Deficiency; GSD; PKU; Metylmalonic Acidemia; フェニルケトン尿症; 変異検出法; STR; 保因者診断; 非ケトーシス型高グリシン血症; 遺伝病; 遺伝子変異; short tandem repeat; 多型診断; アレル特異的PCR

Present study carried out the following projects ;1. Cloning of holocarboxylase synthetase (HCS) geneDeficiency of HCS causes biotin-responsive multiple carboxylase. We have choned the human HCScDNA,which shows homology to BirA and maps to chromosome 21q22.1.2. Mutation studies from various genetic disorders :1) Mutation analysis in 6 Japanese patients revealed the misssense mutation of T997C,G1935A and one base deletion (G1067). These mutations together account for about 83% of Japanese HCS alleles. 2) Mutations of GSD patients ; R83H,P257L,R170X,g727t and IVS 1nt-a, together account for 98% of Japanese GSD mutant alleles.The g727t splicing mutation was the most common mutation among the Japanese patients. 3) Mutaions of PKU patients ; R413P,IVS-4nt-1, R111X,Y204C,R243Q,R252W,R241C,R278I and IVS-9, account for 67% of Japanese PKU mutant alleles. 4) Mutaions of PKU patients ; R243Q,R413P,IVS-4, R365X,R111X,R261Q,Y204C,account for 61% of Chinese PKU mutant alleles. 5) Mutations of Japanese methylmalonic acidemia ; G425T and 769DELTACA.6) Mutations of Japanese CPT II deficient Patients ; F352C,V368I and M647V.7) Mutation of Japanese familial ALS ; H46R.3. Development of multiplex allele specific PCR ; We have applied ASPCR methodology to the detection of mutations in PAH gene. Single ASPCR tests have been developed for 12 mutations found in Japan and China. ASPCR reactions for 3 or 4 mutaions have been multiplexed. The two multiplex tests will detect the presence of the R111X,R261Q and Y204C mutations and the R243Q,R413P,IVS 4nt-1 and Y356X mutations, respectively.

本研究进行了以下项目； 1。 HCS的全羧化酶合成酶（HCS）的克隆引起生物素反应性多重羧化酶。我们cho了人类的HCSCDNA，该HCSCDNA与Bira显示同源，并向21q22.1.2染色体显示。来自各种遗传疾病的突变研究：1）6名日本患者的突变分析显示，T997C，G1935A和一种基本缺失（G1067）的Missensensense突变（G1067）。这些突变共同占日本HCS等位基因的83％。 2）GSD患者的突变； R83H，P257L，R170X，G727T和IVS 1NT-A，占日本GSD突变等位基因的98％。G727T剪接突变是日本患者中最常见的突变。 3）PKU患者的突变； R413P，IVS-4NT-1，R111X，Y204C，R243Q，R252W，R241C，R278I和IVS-9，占日本PKU突变等位基因的67％。 4）PKU患者的突变； R243Q，R413P，IVS-4，R365X，R111X，R261Q，Y204C，占中国PKU突变等位基因的61％。 5）日本甲基甲酸酸血症的突变； G425T和769Deltaca.6）日本CPT II缺乏的突变； F352C，V368I和M647V.7）日本家族性ALS突变； H46r.3。多重等位基因特异性PCR的发展；我们已将ASPCR方法应用于PAH基因中突变的检测。在日本和中国发现的12个突变已经开发了单个ASPCR测试。 3或4个突变的ASPCR反应已被多路复用。这两个多路复用测试将分别检测R111X，R261Q和Y204C突变以及R243Q，R413P，IVS 4NT-1和Y356X突变的存在。

项目成果

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Ikeda, H.等人：“二氢蝶啶还原酶缺乏症的分子分析：日本患者中两种新突变的鉴定。”

Suzuki,Y.et al.: "Enzymatic diagnosis of holocarboxylase synthetase deficiency using apocarboxyl carrier protein as a substrate." Clinica Chemica Acta. 251. 41-52 (1996)

Suzuki,Y.et al.：“使用脱辅基羧基载体蛋白作为底物对全羧化酶合成酶缺陷进行酶法诊断。”

Suzuki,Y.et al.: "Isolation and characterization of mutations human holocarboxylase synthetase cDNA." Nature Genetics. 8. 122-128 (1994)

Suzuki,Y.et al.：“人全羧化酶合成酶 cDNA 突变的分离和表征。”

Suzuki.Y.et al: "Enzymatic diagnosis of holocarboxylase synthetase deficiency using apo-carboxyl carrler protein as a substrate." Clinica Chemica Acta. 251. 41-52 (1996)

Suzuki.Y.et al：“使用脱辅基羧基 carrler 蛋白作为底物对全羧化酶合成酶缺陷进行酶法诊断。”

Aoki,Y.et al.: "Characterization of mutant holocarboxylase synthetase (IICS) : a Km was not elevated in a patient with HCS deficiency." Pediatric Research. (in press). (1997)

Aoki,Y.et al.：“突变型全羧化酶合成酶 (IICS) 的表征：HCS 缺乏症患者的 Km 没有升高。”