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Method and kit for detecting susceptibility of ankylosing spondylitis

An in vitro detection and specific technology, applied in the fields of molecular biology and medicine, can solve the unconfirmed reports of the correlation between TAP1 gene and ankylosing spondylitis, the unconfirmed reports of the correlation between TAP1 gene SNP and ankylosing spondylitis, etc. question

Inactive Publication Date: 2009-09-09
CHINESE NAT HUMAN GENOME CENT AT SHANGHAI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0010] Although there have been many studies on various gene polymorphisms and ankylosing spondylitis, there is no report confirming the correlation between TAP1 gene and ankylosing spondylitis, and even There is no report confirming the correlation between the TAP1 gene SNP described in the present invention and ankylosing spondylitis

Method used

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  • Method and kit for detecting susceptibility of ankylosing spondylitis
  • Method and kit for detecting susceptibility of ankylosing spondylitis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0059] 1.1 Research object

[0060] All case samples were collected from outpatients, and the diagnosis of the patients was made by rheumatologists with rich clinical experience in Renji Hospital according to the revised New York criteria proposed in 1984, and the diagnosis was confirmed through multiple follow-up visits. Control samples were selected from age- and sex-matched individuals with no history of arthritis in the Southern Central Sample Bank.

[0061] Peripheral blood samples were randomly collected from 192 patients and 463 normal controls on the basis of informed consent.

[0062] 1.2 Experimental methods and results

[0063] 1.2.1 DNA extraction

[0064] DNA was extracted from human peripheral blood samples by the conventional phenol-chloroform method, and the concentration was corrected to 20ng / ul for conventional PCR amplification.

[0065] 1.2.2 Design of PCR and sequencing primers

[0066] According to the genome sequence of TAP1 in GenBank, the following...

Embodiment 2

[0080] Ankylosing spondylitis susceptibility detection kit

[0081] As described in Example 1, the G→T mutation at position 430 of SEQ ID NO: 1 is closely related to ankylosing spondylitis. Therefore, specific primers for the TAP1 gene can be designed based on this mutation and then amplified using the patient's DNA as a template for detection.

[0082] Prepare a test kit (100 person-times), which contains:

[0083]

[0084] A test group composed of 100 individuals was randomly selected, including subjects whose ankylosing spondylitis was unknown, patients known to have ankylosing spondylitis, and normal subjects who were tested to be free of ankylosing spondylitis.

[0085] Take 3ml of peripheral blood from the subject to be tested in the test group, and use conventional methods (or use a specific kit) to extract DNA from the blood. Dilute the PCR primers in the Ankylosing Spondylitis Detection Kit to lìmol / ìl, and use the extracted DNA as a template to perform a PCR react...

Embodiment 3

[0091] Auxiliary testing for susceptibility to ankylosing spondylitis

[0092] The test in Example 2 was repeated, with the difference that 75 people (who did not know whether they had symptoms of ankylosing spondylitis before the test) were randomly selected for testing.

[0093] Prepare a test kit (100 person-times), which contains:

[0094]

[0095] Take 3ml of peripheral blood from the subject to be tested, and use conventional methods (or use a specific kit) to extract DNA from the blood. Dilute the PCR primers in the Ankylosing Spondylitis Detection Kit to 1μmol / μl, and use the extracted DNA as a template to perform a PCR reaction with the provided primers. After the PCR product was purified, it was sequenced with an ABI 3730 DNA sequencer, and the sequence was read and SNP confirmed with Polyphred software.

[0096] The results also confirmed that the proportion of ankylosing spondylitis in the test subjects containing 430 G→T was significantly higher than that of ...

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Abstract

The invention discloses a method for detecting susceptibility of ankylosing spondylitis, which comprises the step of detecting whether transporter antigenic peptides 1 (TPA1), transcripts and / or protein of an individual exist mutation or not compared with normal TPA1, normal transcripts and / or normal protein; and if the mutation exists, the possibility that the individual suffers from the ankylosing spondylitis is larger than the possibility that general group suffers form the ankylosing spondylitis. The invention also discloses a corresponding detecting kit.

Description

technical field [0001] The present invention relates to the fields of molecular biology and medicine. More specifically, it relates to the single nucleotide polymorphism (singlenucleotide polymorphism, SNP) of antigen peptide transporter 1 (Transporter of Antigen Peptides 1, TAP1) and its correlation with ankylosing spondylitis. The present invention also relates to methods and kits for detecting these SNPs. Background technique [0002] Ankylosing spondylitis, also known as Von Bechterev's disease or Maritstrumpell's disease, is a chronic, progressive, chronic inflammatory disease involving axial joints. It mainly affects the sacroiliac joints, spinal joints and paravertebral tissues of the pelvis. The main symptoms were low back pain, spinal stiffness and limited range of motion, and X-ray showed sacroilitis on both sides. Since the disease generally invades the sacroiliac joints first, and then mainly involves the spine, eventually leading to bony ankylosis of the spin...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 黄薇雷蓉牛振民
Owner CHINESE NAT HUMAN GENOME CENT AT SHANGHAI
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