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 high myopia causative genes and other problems

Active Publication Date: 2011-11-23
SICHUAN ACADEMY OF MEDICAL SCI SICHUAN PROVINCIAL PEOPLES HOSPITAL
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  • 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 cases collected by the National Natural Science Foundation of China (project number: 30900809, 30871350) undertaken by the Sichuan Provincial Key Laboratory of Human Disease Gene Research, Sichuan Academy of Medical Sciences Sichuan Provincial People's Hospital. A total of 697 cases of high myopia patients were collected, with an average age of 39.7±11.3 years, of which 38.6% were male, and 1086 cases of controls (normal controls, the inclusion criteria were: over 40 years old, long-term close work, diopter>-1.0D.), The mean age was 62.7±10.2 years old, and 37.7% of them were male. All subjects were of Han nationality and signed informed consent, and this study was also approved by the ethics committee.

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

Embodiment 2

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

[0046] The present invention adopts the Snapshot genotyping technology to detect five sites of rs9510902, rs9318086, rs3794338, rs7325450 and rs7331047.

[0047] The experimental steps are as follows:

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

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

[0050] 3. Snapshot reaction: Snapshot MULTIPLEX 1μl, purified PCR mi...

Embodiment 3

[0101] Correlation between embodiment 3 SNP loci and high myopia

[0102] Statistical methods: SPSS13.0 software was used for analysis, and the Hardy-Weinberg equilibrium test was tested. Quantitative variables were tested by t test, and quality variables were tested by chi-square test. Two-sided P<0.05 was considered statistically significant. The result is as follows:

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

[0104]

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

[0106] 2) SNP polymorphisms are closely related to high myopia

[0107] rs9510902:

[0108]

[0109] Without considering the traditional risk factors of 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 its 95% CI is 1.19>1, and it can be determined that the A allele of rs9510902 is a risk factor; the homozygous carrier (AA) will increase the risk of the disease by 1.87 times...

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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 a reagent for predicting susceptibility to high myopia, more specifically by measuring the single nucleotide polymorphism (single nucleotide polymorphism, SNP) sites rs9510902(G→A), rs9318086( G→A), rs3794338(C→T), rs7325450(A→G) and rs7331047(T→C) predict the susceptibility of subjects to high myopia. This method can be used for auxiliary diagnosis, treatment and new drug development of diseases. Belongs to the field of biotechnology. Background technique [0002] According to the degree of myopia, myopia is usually divided into three categories: ① less than 3.00D (300 degrees) is called mild myopia; ② 3.00D ~ 6.00D (300 ~ 600 degrees) is moderate myopia; ③ 6. Those above 00D (600 degrees) are highly myopic. [0003] High myopia is usually accompanied by elongation of the eye axis and deformation of the posterior pole of the eye, including pathological changes such as sclera thinning and choroidal atroph...

Claims

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Application Information

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Patent Type & Authority Applications(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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