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Kit and method for determining cfhr1 genotype, cfhr1 on high-density lipoprotein and oxidized phosphorylcholine

A high-density lipoprotein and phosphorylcholine technology, which is applied in biochemical equipment and methods, microbial measurement/testing, biological testing, etc., can solve problems such as the unknown risk mechanism of AMD

Inactive Publication Date: 2018-06-08
张康
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the mechanism by which it affects AMD risk is largely unknown

Method used

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  • Kit and method for determining cfhr1 genotype, cfhr1 on high-density lipoprotein and oxidized phosphorylcholine
  • Kit and method for determining cfhr1 genotype, cfhr1 on high-density lipoprotein and oxidized phosphorylcholine
  • Kit and method for determining cfhr1 genotype, cfhr1 on high-density lipoprotein and oxidized phosphorylcholine

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Experimental program
Comparison scheme
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Embodiment 1

[0133] Composition and use of embodiment one kit of the present invention

[0134] 1. Use of genotyping kit

[0135] a. DNA template extraction from human tissue samples

[0136] b. Preparation of peripheral blood genomic DNA as a template for determination;

[0137] The method for the preparation of the assay template is:

[0138] 10XNEB Buffer 1ul

[0139] dNTP(10uM) 0.25ul

[0140] Primer-F(10uM) 0.25ul

[0141] Primer-R (10uM) 0.25ul

[0142] NEB Taq (5U / ul) 0.05ul

[0143]DNA (25-50ng / ul) 1-1.5ul

[0144] H2O supplemented to 10ul

[0145] PCR: 95 degrees for 5 minutes

[0146] 95 degrees 30s

[0147] 58 degrees 30s,

[0148] 72 degrees 30s 30-32cycle

[0149] 72 degrees 5min

[0150] Electrophoresis to verify the success of PCR.

[0151] The primer prepared by DNA template among the present invention is:

[0152] rs6677604-tagged CFHR1

[0153] Upstream primer: 5'-GATGGAAGACACCAGAGCAGAT-3' (SEQ ID No 1)

[0154] Downstream primer: 5'-GAATCCATCAGCTTCTACCAT...

Embodiment 2

[0212] Embodiment 2 Pathological experiments and results related to human AMD

[0213] 1. Immunohistochemistry of Human AMD Eyes

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Abstract

The technical problem to be solved by the present invention is to provide diagnostic reagents for the occurrence and development of age-related macular degeneration (AMD). The invention relates to a kit for genotype identification of CFHR1, determination of CFHR1 content in plasma samples, CFHR1 binding amount on high-density lipoprotein-like particles, and CFHR1 binding to oxidation-specific antigen and a detection method thereof. The invention provides a reliable reference index for the diagnosis of the occurrence and development of age-related macular degeneration (AMD). Creatively identified different genotypes of CFHR1. And the work of the present invention may lead to new strategies in the prevention and treatment of senile macular degeneration, such as through the inhibition of CFHR1 activity or its downstream participation in choroidal formation inflammatory genes, including the inhibition of IL-1 activity, in order to prevent and treat senile macular degeneration macular degeneration. It has broad development and application prospects.

Description

technical field [0001] The present invention relates to the genotype identification of complement factor H-related protein 1 (CFHR1), the determination method of CFHR1 in plasma, the separation and purification of high-density lipoprotein-like particles containing CFHR1, the binding of CFHR1 to oxidation-specific antigens, and high-density lipoprotein Detection of oxidized phospholipids on protein (HDL), and through the data obtained from the above detection, early warning diagnosis and treatment guidance for age-related macular degeneration. It specifically relates to a relevant quantitative detection kit and a detection method thereof. Background technique [0002] The pathogenesis of age-related macular degeneration (AMD) involves the interaction of individual genetic variation and environmental risk factors such as oxidative stress. Severe loss of vision in AMD patients is mostly due to choroidal neovascularization (CNV) hemorrhage and exudation, or wet AMD. AMD is the...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/11C12Q1/6883G01N33/68G01N33/53G01N33/543
Inventor 张康彼得.肖
Owner 张康