Susceptibility gene region closely related to psoriasis and method for detecting psoriasis susceptible population

A susceptibility gene, susceptible population technology, applied in the fields of molecular genetics and dermatology, can solve problems such as lack of identification methods

Inactive Publication Date: 2009-09-09
安徽医科大学皮肤病研究所
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For how to identify genetic susceptibility genes and the genetic susceptibility of test subjects, there has been a lack of comprehensive, systematic and effective identification methods in this field

Method used

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  • Susceptibility gene region closely related to psoriasis and method for detecting psoriasis susceptible population
  • Susceptibility gene region closely related to psoriasis and method for detecting psoriasis susceptible population
  • Susceptibility gene region closely related to psoriasis and method for detecting psoriasis susceptible population

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] The following uses the MassARRAY system (sequenom, Inc) to detect the LCE gene SNP difference in this Block region and the haplotype difference derived therefrom to determine the susceptible population of psoriasis.

[0031] 1. Extraction of genomic DNA

[0032]The subjects all draw 5ml of anticoagulant blood from the peripheral vein, and use the DNA extraction kit to extract genomic DNA. The steps are as follows: ① Pipette 1.0ml cell lysate (FG1) into a labeled 2.0ml centrifuge tube, take 400ul blood sample into the corresponding centrifuge tube, invert and mix 20-30 times until no blood clots appear, centrifuge at 16,000g 4 minutes. ②Carefully pour the supernatant, drain on absorbent paper for 30 seconds, add 200ul of denaturation buffer / proteinase K (FG2 / PK) solution, shake and mix each part immediately until no cell clumps appear, wait until all of them are added. Check carefully for incompletely lysed samples. If so, it should be shaken thoroughly. ③After a short centr...

Embodiment 2

[0073] Next, the differences in the SNP sites of the LCE gene in this Block region are detected by gene sequencing to determine the susceptible population of psoriasis.

[0074] 1. Extraction of genomic DNA

[0075] After extracting anticoagulant blood from the peripheral vein of the subject, genomic DNA is extracted. The steps are shown in the first part of the first implementation method.

[0076] 2. PCR amplification of target fragments containing SNP sites

[0077] Design the upstream and downstream primers that can amplify the SNP site of the LCE gene in this Block region, and configure it as follows

[0078]

Reagent

Concentration

Volume

(μL)

HPLC grade water

N / A

1.750

10X PCR buffer containing 15mM MgCl2

1.25x(1.875mM MgCl2)

0.625

25mM MgCl2

1.625mM

0.325

25mM dNTP mix

500uM

0.100

0.5uM primer

0.1uM

1.000

5U / μl HotStar DNA polymerase

1Unit

0.200

...

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Abstract

The invention relates to a susceptibility gene region closely related to psoriasis and a method for detecting psoriasis susceptible population. The susceptibility gene region contains 7 genes, that is, LCE3A, LCE3B, LCE3C, LCE3D, LCE3E, LCE2C and LCE2D. The method for detecting psoriasis susceptible population by the region is realized by the following steps: extracting genome DNA; amplifying PCR; purifying by SAP alkaline phosphatase; allowing single base extension reaction of SNP sites according to a UEP primer designed by the SNP sites in the region; scanning by a mass spectrometer, judging difference of the SNP sites in the susceptibility gene region according to different values of molecular weights of various single base extension products; and judging results. The invention discloses the susceptibility gene region closely related to psoriasis, and provides the method for accurately and effectively determining the psoriasis susceptible population according to hereditary properties of the susceptibility gene region.

Description

Technical field [0001] The invention relates to the fields of molecular genetics and dermatology. Specifically, the present invention relates to a method in which a susceptible gene region closely related to psoriasis has been used to detect people who are susceptible to psoriasis. Background technique [0002] In the field of skin diseases, psoriasis (psoriasis) is a common chronic and recurrent complex disease with erythema and scales as the main manifestations. It was listed by the World Health Organization as one of the two major chronic diseases in the field of human skin diseases in the 20th century. 1. The incidence of this disease is as high as 2% among Caucasians, while it is 0.123% among Chinese population. Approximately 5 million individuals in China suffer from this disease, and it is increasing at a rate of 100,000 cases each year. Psoriasis is commonly known as "psoriasis", which is easy to relapse, difficult to cure, and protracted, and patients are prone to joint ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/12C12Q1/68
Inventor 张学军杨森刘建军张风雨孙良丹
Owner 安徽医科大学皮肤病研究所
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