| Project/Area Number |
14570376
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Public health/Health science
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| Research Institution | NATIONAL CARDIOVASCULAR CENTER RESEARCH INSTITUTE |
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Principal Investigator |
IKEDA Yasuyuki National Cardiovascular Center Research Institute, Etiology and Pathophysiology, Senior staff, 病因部, 室長 (90176107)
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| Co-Investigator(Kenkyū-buntansha) |
TAKAGI Atsuko National Cardiovascular Center Research Institute, Pharmacology, Senior staff, 薬理部, 室長 (90179416)
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| Project Period (FY) |
2002 – 2003
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| Project Status |
Completed (Fiscal Year 2003)
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| Budget Amount *help |
¥2,900,000 (Direct Cost: ¥2,900,000)
Fiscal Year 2003: ¥1,600,000 (Direct Cost: ¥1,600,000)
Fiscal Year 2002: ¥1,300,000 (Direct Cost: ¥1,300,000)
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| Keywords | Lipoprotein lipase / Hypertriglyceridemia / type IV hyperlipoproteinemia / Insulin resistance / Heart disease / Gene analysis / 電気化学的アレイ / 高インシュリン血症 / IV型高脂血症 |
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| Research Abstract |
Subjects (carriers) with heterozygous LPL deficiency are prone to develop mild hypertriglyceridemia (type IV hyperlipoproteinemia) when complicated with environmental factors such as a hyperinsulinemic state and/or a high alcohol intake, which stimulate triglyceride synthesis in the liver, while carriers are normolipidemic provided that they do not have the environmental factors. Identification of heterozygous LPL gene mutations as an early diagnosis, therefore, is important for preventing the development of hypertriglyceridemia and subsequent development of atherosclerosis by getting the patient to change to a more healthful lifestyle. We aimed to develop and evaluate an electrochemical hybridization assay with ferrocenylnaphthalene diimide (FND) for the detection of heterozygous LPL gene mutations.
We developed the electrochemical hybridization assay for the detection of heterozygous LPL mutations, which are heterozygotes for a G-to-A transition at the 818th position (G188E : 818G normal allele and 818A mutant allele) and for a deletion of G at the 916th of the LPL gene-exon 5 (916G normal allele and 916G-del mutant allele). Discriminating between a wild type (Wt) allele and mutant type (Mt) allele is based on the hybridization between two probes such as a Wt probe and a Mt probe (13-15 mer) immobilized on a gold electrode and the PCR product od exon 5 (350 bp). The hybridized double-stranded DNA was quantified by a differential pulse voltammetry at 460 mV using FND as an intercalator.
The electrochemical hybridization assay with FND allowed quick discrimination among a Wt (818G)/Wt (818G) homozygote, a Wt (818G)/Mt (818A) heterozygote and a Mt (818A)/Mt (818A) homozygote of the LPL gene. The same results were obtained for the discrimination of a Wt (916g) allele and a Mt (919G-del) allele. This method is also applicable for the detection of other LPL gene mutations.
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