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Microflow controlled method for detecting subtypes of C 482T polymorphism of apolipoprotein CIII gene

A C-482T, apolipoprotein technology, applied in the field of gene detection, can solve the problems of complexity and large variation of polymorphism ratio, and achieve the effects of simple operation, high resolution and low cost

Inactive Publication Date: 2007-07-25
AFFILIATED HOSPITAL OF NANTONG UNIV
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Problems solved by technology

However, the Apo CIII gene polymorphism is closely related to the origin of different ethnic groups, and the proportion of Apo CIII C-482T gene polymorphism in different populations in different regions varies greatly
At present, there are few domestic reports on the relationship between Apo CIII C-482T gene polymorphism and disease, and there is no research on its relationship with HTG; after amplification of the Apo CIII-482 site, the difference between the two fragments is only 16bp, Usually separated by polyacrylamide gel electrophoresis, but more complicated

Method used

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Embodiment

[0014] 1. Source of material: (1) 74 cases of normal control group were from the Physical Examination Center of the Affiliated Hospital of Nantong University. There were 37 females and 37 males, aged 24 to 68 years (mean age 40±10 years). (2) 112 cases of HTG patients selected TG>2.2mmol / L, CH<6.2mmol / L, liver, kidney, thyroid and other diseases were excluded, including 62 females and 50 males, aged 23-82 years (average age 56±14 years old), all were inpatients in the Affiliated Hospital of Nantong University, and the above patients and the normal control group were all unrelated Han nationality.

[0015] 2. Main instruments and reagents: Caliper 1000 microfluidic chip instrument, microfluidic chip, and ME polymer glue were purchased from Shanghai Haoyuan Biotechnology Co., Ltd. (Agilent, USA), Dolphin-Doc gel automatic imaging system (WEALTEC, U.S.), PTC-200 PCR amplification instrument (MJ, U.S.), U-0080D spectrophotometer (Hitachi, Japan). Restriction enzyme MspI was purc...

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Abstract

This invention discloses a method for detecting C-482T polymorphism of apolipoprotein CIII gene with micro-fluidic chip. The method comprises: genome DNA extraction, nucleic acid amplification, enzyme digestion, and micro-fluidic chip electrophoresis. The method has such advantages as high resolution and easy operation.

Description

Technical field: [0001] The invention relates to a gene detection method. Background technique: [0002] Apolipoprotein III (Apo CIII) is a single polypeptide chain composed of 79 amino acid residues with a molecular weight of 818KD. It is mainly synthesized in the liver and expressed in a small amount in the small intestine. Apo CIII affects the metabolism of plasma triglyceride-rich lipoprotein (TRL) by inhibiting the activity of enzymes and receptors such as lipoprotein lipase (LPL) and hepatic lipase (HL). If the expression of Apo CIII is out of control, the rate of lipolysis of chylomicrons (CM) and very low-density lipoprotein (VLDL) will be delayed, thus causing hypertriglyceridemia (HTG). A large number of epidemiological investigations have shown that triglyceride (TG) is an independent risk factor for coronary heart disease (CHD) and atherosclerosis (AS). The Apo CIII gene is located at 11q23, has 4 exons and 3 introns, and is closely linked with the Apo AI and A...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 王惠民张志泉周锦红俞娟陈莲英吴翔
Owner AFFILIATED HOSPITAL OF NANTONG UNIV
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