Method and kit for detecting susceptibility of ankylosing spondylitis

A technique for ankylosing spondylitis, kits, applied in the fields of molecular biology and medicine

Inactive Publication Date: 2013-03-27
上海市生物医药技术研究院
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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

[0060] 1.1 Research object

[0061] 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.

[0062] Peripheral blood samples were randomly collected from 195 patients and 471 normal controls on the basis of informed consent.

[0063] 1.2 Experimental methods and results

[0064] 1.2.1 DNA extraction

[0065] 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.

[0066] 1.2.2 Design of PCR and sequencing primers

[0067] According to the genome sequence of TAP1 in GenBank, the following...

Embodiment 2

[0081] Ankylosing spondylitis susceptibility detection kit

[0082] As described in Example 1, the mutation at position 621 G→T in 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.

[0083] Prepare a test kit (100 person-times), which contains:

[0084] name sequence concentration sense primer SEQ ID NO: 2 100pmol antisense primer SEQ ID NO: 3 100pmol PCR reaction solution Containing Taq Enzyme dNTP Magnesium Ion PCR Reaction Buffer

[0085] 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.

[0086] Take 3ml of peripheral blood from the subject to be tested in the test group, ...

Embodiment 3

[0092] Auxiliary testing for susceptibility to ankylosing spondylitis

[0093] The test in Example 2 was repeated, with the difference that 80 people (who did not know whether they had symptoms of ankylosing spondylitis before the test) were randomly selected for testing.

[0094] Prepare a test kit (100 person-times), which contains:

[0095] name sequence concentration sense primer SEQ ID NO: 2 100pmol antisense primer SEQ ID NO: 3 100pmol PCR reaction solution Containing Taq Enzyme dNTP Magnesium Ion PCR Reaction Buffer

[0096] 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...

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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. [0002] Background technique [0003] 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...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68
Inventor 黄薇雷蓉牛振民
Owner 上海市生物医药技术研究院
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