Method and reagent for prediction of ankylosing spondylitis susceptibility

A technology for ankylosing spondylitis and susceptibility, which can be used in anti-inflammatory agents, non-central analgesics, biochemical equipment and methods, etc., and can solve problems such as no AIF1 gene research results

Inactive Publication Date: 2013-07-17
卫生部北京医院
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] At present, there are no reports on the association between AIF1 gene and autoimmu

Method used

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  • Method and reagent for prediction of ankylosing spondylitis susceptibility
  • Method and reagent for prediction of ankylosing spondylitis susceptibility
  • Method and reagent for prediction of ankylosing spondylitis susceptibility

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Example 1: Blood Sample Collection and Genomic DNA Extraction

[0047] The selected cases were selected according to the revised diagnostic criteria in New York in 1984. A total of 118 unrelated AS patients (age: 14-44 years old, average 24 years old) from the 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. .

[0048] Human genomic DNA was prepared according to the following method. ①First 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 stand at room temperature for 10 minutes; ②C...

Embodiment 2

[0049] Embodiment 2 Identification and identification of SNP

[0050] The present invention adopts PCR-high resolution melting curve (HRM) analysis method and PCR sequencing technology to simultaneously carry out the genotype of the +879 site (its allelic site is C / T) of the 2nd intron region of AIF1 gene detection. figure 2 It is the genotyping map of AIF1 gene variation site by HRM method, image 3 Sequencing map of AIF1 gene mutation site.

[0051] 1) Determination of PCR-HRM primers

[0052] The DNA base sequence (SEQ ID No.1) near +879 was retrieved from Genebank, and the primer design was completed under Oligo7.0 software. The target fragment is located in the second intron region of the AIF1 gene, with a total length of 58bp. The sense strand F1 (+848bp-+867bp) and the antisense strand R1 (+887bp-+905bp) are determined. The specific primer sequences are as follows:

[0053] F1: 5'-GGTGTGCAGGACTAAGAAGA-3' (SEQ ID No. 2)

[0054] R1: 5'-CAAATCGTGAGGAATGGAG-3' (SEQ I...

Embodiment 3 Embodiment 3

[0063] Example 3 Example 3 Correlation between gene SNP and AS

[0064] 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 alleles and genotypes of the AIF1 gene +879 polymorphic site between the AS case group and the normal control group, and the AS risk OR value and its 95 %CI confidence interval, with P<0.05 as the standard of significant difference.

[0065] Results: The distribution of the genotype and allele frequency of the SNP+879 polymorphic site on the AIF1 gene located in the 6p21.3 region between the cases and the control group is shown in Table 2.

[0066] Table 2 Distribution of the genotype and allele frequency of the AIF1 gene +879C / T polymorphic site in the case-control group

[0067]

[0068] Note: OR: odds ratio; CI: confidence interval. *T allele is the risk allele for AS. Subjects were ...

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Abstract

The invention discloses a method and a reagent for prediction of ankylosing spondylitis susceptibility. According to the invention, genomic DNA of a host cell is extracted and the genotype of site + 879 in a second intron zone of the AIF1 gene of a subject is determined so as to predicate susceptibility of the subject to ankylosing spondylitis; if the genotype of the single nucleotide polymorphism site + 879 is CC, lowest ankylosing spondylitis susceptibility is obtained, and if the site + 879 contains a T allele, i.e., the genotype is CT or TT, highest susceptibility is obtained. The invention has the following advantages: correlation between the AIF1 gene polymorphism site and the ankylosing spondylitis is elaborated for the first time; the method for predicting ankylosing spondylitis susceptibility is provided; and the method is applicable to prevention, auxiliary diagnosis and treatment of ankylosing spondylitis and can also be used for research and development of a novel drug.

Description

technical field [0001] The present invention relates to a method and reagent for predicting the susceptibility of ankylosing spondylitis, more specifically, predicting the susceptibility of subjects to ankylosing spondylitis by determining the polymorphism of gene AIF1 related to ankylosing spondylitis. The method can be used for auxiliary diagnosis of diseases and development of new medicines, and belongs 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 a...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11A61K48/00A61P29/00A61P19/00
Inventor 杨泽杨帆朱小泉周林史晓红唐雷孙亮
Owner 卫生部北京医院
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