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Screening kit for detecting high myopia

A high myopia, kit technology, applied in the determination/inspection of microorganisms, biochemical equipment and methods, etc., can solve the problem of not finding the high myopia causative gene and so on

Active Publication Date: 2013-05-22
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, no definite high myopia-causing gene has been found so far

Method used

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  • Screening kit for detecting high myopia
  • Screening kit for detecting high myopia
  • Screening kit for detecting high myopia

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Example 1 Sample collection and extraction of genomic DNA

[0043] The samples are collected by the National Natural Science Foundation of China (project number: 30900809, 30871350) undertaken by the Sichuan Provincial Key Laboratory of Human Disease Genetic Research, Sichuan Academy of Medical Sciences and Sichuan Provincial People's Hospital. A total of 697 patients with high myopia, with an average age of 39.7±11.3 years old, were collected, of which 38.6% were males, and 1086 controls (normal controls, the selection criteria are: over 40 years old, long-term close work, diopter> -1.0D.), The average age is 62.7±10.2 years, of which 37.7% are males. All subjects were Han nationality and signed an informed consent form. This study was also approved by the ethics committee.

[0044] According to the following method, genomic DNA was prepared from human peripheral blood. In the presence of the anticoagulant EDTA, the collected 5 ml of human peripheral blood was centrifuged...

Embodiment 2

[0045] Example 2 PCR amplification and genotype analysis of DNA fragments containing rs9510902, rs9318086, rs3794338, rs7325450 and rs7331047 single nucleotide polymorphisms

[0046] The present invention uses Snapshot genotyping technology to detect five loci rs9510902, rs9318086, rs3794338, rs7325450 and rs7331047.

[0047] The experimental steps are as follows:

[0048] 1. PCR: Denaturation at 95°C for 5 minutes; 95°C for 30 seconds, 55°C annealing for 30 seconds, 72°C extension for 45 seconds, 35 cycles; 72°C extension for 7 minutes, 4°C incubation.

[0049] 2. Purification 1: Take 1-3μl each of the PCR product from step 1 (depending on the number of PCRs to be mixed, the total amount is 9μl, add 3μl of the mixture (SAP 2μl, 1:10 Exol enzyme 1μl). 1:10Exol enzyme Preparation: 1 volume of Exol enzyme, 1 volume of SAP 10Xbuffer, 8 volume of ddH 2 O. Procedure: 37°C for 1h, 75°C for 15min.

[0050] 3. Snapshot reaction: Snapshot MULTIPLEX 1μl, purified PCR mix 1μl, Snapshot primer 0....

Embodiment 3

[0101] Example 3 The correlation between SNP sites and high myopia

[0102] Statistical method: use SPSS13.0 software to analyze, test Hardy-Weinberg balance test, use t test for quantity variables, and use chi-square test for quality variables. Two-sided P<0.05 was considered statistically significant. The results are as follows:

[0103] 1) Basic clinical data of cases and controls

[0104]

[0105] Note: **, P<0.01.

[0106] 2) SNP locus polymorphism is closely related to high myopia

[0107] rs9510902:

[0108]

[0109] Without considering the traditional risk factors for high myopia, the A allele can significantly increase the risk of high myopia (OR=1.36, 95%CI=1.19-1.55), that is, the risk of high myopia increases by 1.36 times, and The lower limit of 95% CI is 1.19>1, which can confirm that the A allele at rs9510902 is a risk factor; among them, homozygous carriers (AA) increase the risk of disease by 1.87 times, and heterozygotes (AG) increase the risk of disease by 1.19 Tim...

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Abstract

The invention discloses a screening kit for detecting high myopia, which comprises optional relevant reagents for detecting MIPEP gene rs9510902 mutation, rs9318086 mutation, rs3794338 mutation, rs7325450 mutation, or / and rs7331047 mutation which are used for high myopia screening; the susceptibility of a subject to high myopia is predicted by determining whether mutation exists at the following five SNP sites: rs9510902, rs9318086, rs3794338, rs7325450, and rs7331047. The advantages of the invention are that: the correlation between the above five SNP sites and high myopia is elucidated for the first time; the prediction method for susceptibility to high myopia is simple, is applicable to the prevention, assistant diagnosis and treatment of high myopia, and is also applicable to new drugdevelopment.

Description

Technical field [0001] The present invention relates to a method and reagents for predicting the susceptibility to high myopia, more specifically by measuring the single nucleotide polymorphism (SNP) sites of MIPEP gene rs9510902 (G→A), rs9318086 ( G→A), rs3794338(C→T), rs7325450(A→G), and rs7331047(T→C) predict the subject’s susceptibility to high myopia. This method can be used for disease diagnosis, treatment and new drug development. It belongs to the field of biotechnology. Background technique [0002] According to the degree of myopia, myopia is usually divided into three categories: ① those within 3.00D (300 degrees) are called mild myopia; ② those with 3.00D to 6.00D (300 degrees to 600 degrees) are moderate myopia; ③6. Those above 00D (600 degrees) are highly myopic. [0003] High myopia is usually accompanied by axial lengthening and deformation changes in the posterior pole of the eye, including pathological changes such as sclera thinning, choroidal atrophy and thinn...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68
Inventor 杨正林石毅鲁芳刘小琦张丁丁马誓林鹤成静林婴
Owner SICHUAN ACADEMY OF MEDICAL SCI SICHUAN PROVINCIAL PEOPLES HOSPITAL
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