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Reagent kit for detecting senility macular degeneration disease

A technology for degenerative diseases and age-related macular degeneration, which is applied in the field of kits for detecting HTRA1 gene mutations related to age-related macular degeneration, can solve the problems of lack of monitoring and diagnosis indicators, and achieve the effects of reducing visual damage, improving accuracy, and high correlation

Active Publication Date: 2008-05-07
SICHUAN ACADEMY OF MEDICAL SCI SICHUAN PROVINCIAL PEOPLES HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in the absence of monitoring and diagnostic indicators in the early stage of the disease, it is possible to screen and discover AMD high-risk groups early, and implement early detection and prevention to reduce their visual impairment

Method used

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  • Reagent kit for detecting senility macular degeneration disease
  • Reagent kit for detecting senility macular degeneration disease
  • Reagent kit for detecting senility macular degeneration disease

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] [Example 1] Blood sample collection

[0027] A total of 581 age-related macular degeneration (AMD) patients and 309 controls were selected. The patients were clinically confirmed AMD, including 392 patients with wet AMD and 189 patients with dry AMD. The controls were people who were older than or equal to 60 years old, had no drusen and retinal pigment changes, and matched the age group of AMD patients.

[0028] For all participants, a detailed investigation of their medical and family histories, and a physical examination and detailed ophthalmology specialist examination were performed. After signing the informed consent form, 5-10ml blood samples were collected from each person.

Embodiment 2

[0029] [Example 2] PCR amplification and genotype analysis comprising the rs11200638 gene

[0030] 1. Extract the blood genome DNA of AMD patients and control groups.

[0031]2. PCR amplification of genomic DNA fragments (including rs11200638)

[0032] Primer sequence: Forward: 5'-ATGCCACCCACAACAACTTT-3'

[0033] Reverse: 5'-CGCGTCCTTCAAACTAATCC-3'

[0034] Reaction conditions: 95°C 3minutes, (94°C 30seconds, 52°C 30seconds, 72°C 45seconds)*35cycles

[0035] 3. Genotyping

[0036] Restriction enzyme identification:

[0037] (1) The PCR product was digested with EagI to identify the G allele

[0038] Using NEB restriction enzyme EagI (5'...C^G G C C G...3'), the reaction system is as follows: 3.9ul H 2 O+5ul PCR product+1ul buffer 3+0.1ul EagI; the reaction conditions are as follows: 37°C for 12 hours.

[0039] (2) 2% agarose gel electrophoresis digestion product, and judge the genotype according to the electrophoresis band (AA homozygote is a DNA band of the same size ...

Embodiment 3

[0100] [Example 3] PCR amplification and genotype analysis comprising the rs10490924 gene

[0101] 1. Extract the blood genome DNA of AMD patients and controls.

[0102] 2. PCR amplification of genomic DNA fragments (including rs10490924).

[0103] Primer sequence: Forward: 5'-TACCCAGGACCGATGGTAAC-3';

[0104] Reverse: 5'-GAGGAAGGCTGAATTGCCTA-3'.

[0105] Reaction conditions: 95°C 3minutes, (94°C 30seconds, 52°C 30seconds, 72°C 45seconds)*35cycles

[0106] 3. Genotyping

[0107] Restriction enzyme identification:

[0108] (1) The PCR product was digested with PVUII to identify the G allele.

[0109] Using NEB restriction enzyme PVUII (5'...C A G^C T G...3'), the reaction system is as follows: 3.9ul H 2 O+5ul PCR product+1ul buffer 2+0.1ul PVUII; the reaction conditions are as follows: 37°C for 12 hours.

[0110](2) 2% agarose gel electrophoresis digestion product, and judge the genotype according to the electrophoresis band (TT homozygote is a DNA band of the same size...

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Abstract

The invention provides a kit for detecting age-related macular degeneration disease, which can be used for early monitoring and detection of AMD high-risk groups, implement early prevention and treatment, and reduce their visual impairment. The kit of the present invention targets the 512G→A mutation of the HTRA1 gene and its related sites / genes, has a high correlation with AMD, and cooperates with the analysis of the CHF gene mutation to improve the accuracy of detection.

Description

technical field [0001] The invention relates to a kit for detecting age-related macular degeneration, in particular a kit for detecting HTRA1 gene variation related to age-related macular degeneration. Background technique [0002] Macular degeneration is considered to be a heterogeneous disease characterized by progressive central vision loss and macular retinal pigment epithelial degeneration, and is an important blinding eye disease. Macular degeneration includes age-related macular degeneration and macular dystrophy (stargart disease, best disease, sorsby fundus dystrophy, etc.). Age-related macular degeneration (AMD) has become one of the most important blinding eye diseases for the elderly, with 10 million patients worldwide, and the prevalence of AMD among people aged 60-69 in my country is 7.77%. Over the age of up to 15.33%. Currently there is no effective treatment for AMD, the main reason is that the pathogenesis is still unclear. [0003] In recent years, studi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68G01N33/563
Inventor 杨正林张康鲁芳
Owner SICHUAN ACADEMY OF MEDICAL SCI SICHUAN PROVINCIAL PEOPLES HOSPITAL
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