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A test kit for detecting vkh syndrome

A kit and reagent technology, applied in the field of kits for detecting VKH syndrome, can solve the problems of rare non-HLA antigen-related reports, and achieve the effect of preventing the development of lesions and correct diagnosis

Active Publication Date: 2016-01-13
THE FIRST AFFILIATED HOSPITAL OF CHONGQING MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, most of the research results on the genetic susceptibility of VKH syndrome focus on the correlation between human leukocyte antigens (HLA-DR4 / DR53, HLA-DR1, HLADRB1*0405, etc.) and the disease, and reports on the correlation between non-HLA antigens and the disease Uncommon, and most reported findings are difficult to replicate in other populations

Method used

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  • A test kit for detecting vkh syndrome
  • A test kit for detecting vkh syndrome
  • A test kit for detecting vkh syndrome

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1 Sample collection and extraction of genomic DNA

[0030] The samples came from the First Affiliated Hospital of Chongqing Medical University. A total of 1,559 VKH patients were collected, including 55.0% males and 45.0% females; 5,640 normal controls, 50.8% males and 49.2% females. All subjects were Han nationality, and voluntarily signed the informed consent, and this study was also approved by the ethics committee.

[0031] Human peripheral blood genomic DNA was prepared according to the operation steps provided by the QIAGEN QIA & DNA Mini Blood Kit kit (purchased from QIAGEN, Germany).

Embodiment 2

[0032] 实施例2包括rs78377598、rs77258390、rs78597810、rs12568393、rs12561798、rs76436269,[rs117633859、rs12564219、rs12563505、rs34017352、rs114661385、rs117301158、rs115009271、rs76091599、rs78865162、rs78917656、rs6693659、rs12566159、rs117282985];rs3021304[rs114800139];rs442309 PCR Amplification and Genotype Analysis of Single Nucleotide Polymorphic DNA Fragments of and rs224058 [rs224048, rs224052, rs224057, rs10995307, rs10995281, rs7088592, rs224071, rs224031, rs224032, rs224033]

[0033] The present invention uses SequenomMassARRAYSNP detection technology to detect the mutations of rs78377598, rs77258390, rs78597810, rs12568393, rs12561798, rs76436269, rs3021304, rs442309 and rs224058 (as shown below).

[0034] 1) Use SEQIDNO.1~2, SEQIDNO.3~4, SEQIDNO.5~6, SEQIDNO.7~8, SEQIDNO.9~10, SEQIDNO.11~12, SEQIDNO.13~14, SEQIDNO.15~16 1. The primers shown in SEQ ID NO.17-18 carry out target gene fragments containing specific sites;

[0035] PCR amplification: 94°C for 4min; 94°C for 20s, 56°C for 30...

Embodiment 4

[0125] Embodiment 4 detection kit

[0126] All components, content and method of use in the kit of the present invention are as follows:

[0127] PCR amplification reagent (50 servings)

[0128] Wherein, the primer pair A-I are the primers shown in SEQ ID NO.1-2-SEQ ID NO.19-20 respectively. Primers A-I are PCR amplification primers, and a-i are specific extension primers.

[0129]

components

concentration

volume

Taq enzyme

4ml

SAP (Shrimp Alkaline Phosphatase)

80ul

SAP Buffer

10×

60ul

Single base extension reaction solution

500ul

PCR primer pair A (rs78377598)

10uM

400ul

PCR primer pair B (rs77258390)

10uM

400ul

PCR primer pair C (rs78597810)

10uM

400ul

PCR primer pair D (rs12568393)

10uM

400ul

PCR primer pair E (rs12561798)

10uM

400ul

...

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Abstract

The invention relates to a kit for detecting VKH (Vogt Koyanagi Harada) syndrome. The kit contains primer pairs of a reagent used for amplifying gene segments at loci of rs78377598, rs77258390, rs78597810, rs12568393, rs12561798, rs76436269, rs3021304, rs442309 and rs224058 and a Sequenom MassARRAY single-base extension primer for specifically detecting idiotypes. The kit is used for predicating the genetic predisposition of a subject for the VKH syndrome by detecting whether the SNP (single nucleotide polymorphism) loci of the subject are varied, and can be used for early screening for the occurrence of the VKH syndrome of Chinese Han population and providing a basis for clinical diagnosis and treatment.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular to a kit for detecting VKH syndrome. Background technique [0002] Vogt-Koyanagi-Harada (VKH) syndrome, also known as Vogt-Koyanagi-Harada syndrome, is an inflammatory disease involving multiple systems throughout the body. The syndrome is characterized by: ①sudden uveitis; ②skin lesions such as eyebrow and hair whitening, baldness, and vitiligo; ③nervous system manifestations such as headache, dizziness, and nausea; ④inner ear symptoms such as tinnitus, deafness, and vertigo. symptom. [0003] VKH syndrome begins with an acute attack, and the course of the disease is prolonged and repeated. There are two clinical types, Vogt-Koyanagi type (VK type) with exudative iridocyclitis and Harada type (H type) with exudative choroiditis. VKH syndrome mostly occurs in people of color, and Chinese, Japanese, Brazilian and American Indians are high-risk groups, and it is more likely to occur in...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6883C12Q2600/106C12Q2600/156
Inventor 杨培增杨正林侯胜平杜利平雷博
Owner THE FIRST AFFILIATED HOSPITAL OF CHONGQING MEDICAL UNIVERSITY
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