Mass screening for ion channel gene abnormality in the patients with long QT syndrome diagnosed by Screening Program for Heart Disease

心脏病筛查项目诊断的长QT综合征患者离子通道基因异常的大规模筛查

• 批准号: 10670739
• 负责人: NOMURA Yuichi
• 金额: $ 1.98万
• 依托单位: Kagoshima University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1998
• 资助国家: 日本
• 起止时间: 1998 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10670739/
• 关键词: long QT; CFLP; gene abnormalities; 逆転写PCR; Kチャンネル; Naチャンネル

Many students are diagnosed as having long QT interval using Screening Program for Heart Disease. If their ion channel gene abnormality(s) are determined, it will be helpful on their medication. Cleavase fragment length polymorphism analysis (CFLP) analysis expresses stable reproducibility and has the possibility of analyzing much longer DNA fragments than those possible by single-strand conformation polymorphism (SSCP), which is usually applied in gene analysis. We examined the possibility of ion channel gene mass screening using the CFLP analysis.At first, CFLP analysis was performed in the study of a family with long QT syndrome. CFLP analysis showed a difference between normal genes and the patients' genes in the part be tween S4 and S5 of the KVLQT1 gene in terms of an increased density of a patient's band whose size was around 50 base. A heterozygous mutation (CCGTGG->CCATGG) was indicated using direct sequencing, and the mutation was confirmed by restriction fragment length polymorphism using the enzyme of Nco I. It was confirmed that one point heterozygous mutation in 835 base pair polymerase chain reaction product was detectable using CFLP analysis. It is suggested that CFLP analysis is a useful method for the screening of unknown mutation(s) in large genes.CFLP analysis was applied in other two families with long QT syndrome or three patients with sporadic long QT interval. However, no abnormalities have been found.Further examinations on other patients are necessary. Combination analysis using CFLP and direct sequencing for whole ion channel gene is starting.

许多学生使用心脏病筛查计划被诊断为QT间期延长。如果确定他们的离子通道基因异常(S)，将有助于他们的药物治疗。裂解酶片段长度多态性分析(CFLP)分析具有稳定的重复性，并且可以分析比通常用于基因分析的单链构象多态性(SSCP)更长的DNA片段。我们研究了使用CFLP分析进行离子通道基因大规模筛选的可能性。首先，对一个长QT综合征家系进行CFLP分析。CFLP分析表明，在KVLQT1基因S4和S5之间的部分，正常基因和患者基因的差异在于患者条带的密度增加，其大小约为50个碱基。用直接测序的方法发现了一个杂合性突变(CCGTGG-&GT；CCATGG)，并用NCO I酶的限制性内切酶片段长度多态性分析证实了该突变。CFLP分析证实，835碱基聚合酶链式反应产物中存在一个点杂合性突变。本研究对另外两个QT间期延长综合征家系和3例散发性QT间期延长患者进行了CFLP分析，结果表明，CFLP分析是一种在大基因中筛查未知突变的有效方法。但未发现异常，需对其他患者作进一步检查。利用CFLP和直接测序对全离子通道基因进行联合分析已经开始。

项目成果

野村裕一, 上村順子, 吉永正夫, 西順一郎, 河野幸春, 安田星一郎: "Cleavase fragment length polymorphism法によるQT延長症候群1家系の遺伝子解析"日本小児循環器学会雑誌. 17(1). 20-25 (2001)

Yuichi Nomura、Junko Uemura、Masao Yoshinaga、Junichiro Nishi、Yukiharu Kono、Seiichiro Yasuda：“使用切割酶片段长度多态性方法对长 QT 综合征 1 家族进行遗传分析”日本儿科心脏病学会杂志 17(1)。 20-25（2001）

Nomura,Yuichi, Kamimura,Junko, Yoshinaga,Masao, Nishi,Jun-ichiro, Kono,Yukiharu, Miyata,Koichiro: "Cleavase fragment length polymorphism analysis in a family with long QT syndrome"Jpn J Ped Card Surg. 17 (1). 20-25 (2001)

Nomura、Yuichi、Kamimura、Junko、Yoshinaga、Masao、Nishi、Jun-ichiro、Kono、Yukiharu、Miyata、Koichiro：“长 QT 综合征家族的裂解酶片段长度多态性分析”Jpn J Ped Card Surg。

野村裕一, 上村順子, 吉永正夫, 西順一郎, 河野幸春, 宮田晃一郎: "Cleavase fragment length polymorphism法によるQT延長症候群1家系の遺伝子解析"日本小児循環器学会雑誌. 17(1). 20-25 (2001)

Yuichi Nomura、Junko Uemura、Masao Yoshinaga、Junichiro Nishi、Yukiharu Kono、Koichiro Miyata：“使用切割酶片段长度多态性方法对长 QT 综合征 1 家族进行遗传分析”日本儿科心脏病学会杂志 17(1)。 20-25（2001）