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Reagent for predicting susceptibility of ankylosing spondylitis

Ankylosing spondylitis, reagent technology, applied in the direction of microbial determination/testing, DNA/RNA fragments, recombinant DNA technology, etc.

Inactive Publication Date: 2015-05-13
BEIJING HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] There are currently no research results on the association of the BAG6 gene with AS

Method used

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  • Reagent for predicting susceptibility of ankylosing spondylitis
  • Reagent for predicting susceptibility of ankylosing spondylitis
  • Reagent for predicting susceptibility of ankylosing spondylitis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Example 1: Collection of blood samples and extraction of genomic DNA:

[0041] 1. Case selection:

[0042] The selected cases were selected according to the revised diagnostic criteria in New York in 1984. A total of 136 unrelated AS patients (age: 14-44 years old, average 24 years old) from Jilin area were selected, and 148 healthy control volunteers (age: 39 years old) from the same area were selected. -72 years, with an average of 46 years). All subjects were of Han nationality and signed written informed consent. This study was approved by the Beijing Hospital of the Ministry of Health and the Ethical Review Committee of the Institute of Gerontology of the Ministry of Health. .

[0043] 2. According to the following method, human genomic DNA was prepared.

[0044] 1. Add 1000 μL of erythrocyte lysate to a labeled 1.5mLEP tube, then add 400 μL of LEDTA anticoagulant blood (invert and mix 3-5 times before adding anticoagulant blood), invert and mix well, and let st...

Embodiment 2

[0052] Identification and identification of embodiment 2 SNP

[0053] The present invention adopts PCR-high resolution melting curve (HRM) analysis method and PCR sequencing technology to simultaneously analyze the site of the 23rd intron region of the 3rd exon region of the BAG6 gene (the allelic site is C / T). genotype detection. figure 2 It is the genotyping map of BAG6 gene variation site by HRM method, image 3 It is the sequencing map of the variation site of BAG6 gene.

[0054] 1. Determination of PCR-HRM primers

[0055] The DNA base sequence (Seq ID №1) near rs201165258 was retrieved from Genebank, and the primer design was completed under Oligo7.0 software. The target fragment is located in the 23rd intron region of the BAG6 gene, with a total length of 51bp. The sense strand F1 (+396bp-+421bp) and the antisense strand R1 (+425bp-+446bp) are determined. The specific primer sequences are as follows:

[0056] F1: 5'-TGTATATCAGACCGGAGCTAAAGAGA-3' (SEQ ID No. 2)

[...

Embodiment 3

[0066] Example 3 Correlation between gene SNP and ankylosing spondylitis (AS)

[0067] 1. Statistical method:

[0068] Statistical methods: The Hardy-Weinberg balance test was used to study the group representativeness of the samples. The Pearson chi-square test in SPSS11.0 software was used to calculate the distribution frequency of the alleles and genotypes of the BAG6 gene rs201165258 site between the AS case group and the normal control group, and the AS risk OR value and its 95% CI were credible Interval, with P<0.05 as the standard of significant difference.

[0069] 2. Results: The distribution of the genotype and allele frequency of the rs201165258 site on the BAG6 gene located in the 6p21.3 region between the cases and the control group is shown in Table 2.

[0070] Table 2: Distribution of genotype and allele frequencies of BAG6 gene rs201165258 between case and control groups

[0071]

[0072] Note: OR: odds ratio; CI: confidence interval. *G allele is the ri...

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Abstract

The invention discloses a reagent for predicting the susceptibility of ankylosing spondylitis, and belongs to the technical field of medical biology. The susceptibility of a subject on the ankylosing spondylitis is predicted by extracting the genome DNA of a host cell and measuring the genotype of the SNP site of the BAG6 gene of the subject. According to the invention, the correlation of the polymorphic site of the BAG 6 gene and the ankylosing spondylitis is firstly expounded, and a method for predicting the susceptibility of the ankylosing spondylitis reagent is provided. The method disclosed by the invention can be used for preventing, accessorily diagnosing and curing the ankylosing spondylitis and can also be used for developing new drugs.

Description

technical field [0001] The invention relates to a reagent for predicting the susceptibility of ankylosing spondylitis, more specifically, predicting the susceptibility of a subject to ankylosing spondylitis by determining the polymorphism of AS-related gene BAG6, and the method can be used for disease diagnosis Auxiliary diagnosis, treatment and new drug development belong to the field of biotechnology. Background technique [0002] Ankylosing spondylitis (AS) is an autoimmune disease that mostly occurs in young adults aged 16-40, with a male to female ratio of about 4 to 10:1. Lesions often first occur in the sacroiliac joints, and a small number of severe cases show rigidity of the entire spine. In addition, some patients have different degrees of extra-spinal lesions such as hip joints, eyes, lungs, cardiovascular, and kidneys. The incidence of AS in the Caucasian population is about 1% to 3%, and the prevalence of AS in my country is about 0.2-0.6%, of which more than ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6883C12Q2600/156
Inventor 杨泽朱小泉杨帆孙亮史晓红唐雷原慧萍张玉荣赵承孝赵帆王娜娜惠娟张政
Owner BEIJING HOSPITAL
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