Method for testing susceptibility of ankylosing spondylitis and kit

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 SNP and ankylosing spondylitis, the unconfirmed reports of the correlation between TAP1 gene 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 for testing susceptibility of ankylosing spondylitis and kit
  • Method for testing susceptibility of ankylosing spondylitis and kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0063] 1.1 Research object

[0064] 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.

[0065] Peripheral blood samples were randomly collected from 197 patients and 474 normal controls on the basis of informed consent.

[0066] 1.2 Experimental methods and results

[0067] 1.2.1 DNA extraction

[0068] 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.

[0069] 1.2.2 Design of PCR and sequencing primers

[0070] According to the genome sequence of TAP1 in GenBank, the following...

Embodiment 2

[0087] Ankylosing spondylitis susceptibility detection kit

[0088] As described in Example 1, the change from (GGG) to (-AA) at position 366 G→deletion in SEQ ID NO: 1 and at positions 185 and 601 in SEQ ID NO: 4 is associated with ankylosing spondylitis Diseases are closely related. Therefore, TAP1 and LMP2 gene-specific primers can be designed based on this mutation to amplify and detect using the patient's DNA as a template.

[0089] Prepare a test kit (100 person-times), which contains:

[0090]

[0091] 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.

[0092] 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 ...

Embodiment 3

[0098] Auxiliary testing for susceptibility to ankylosing spondylitis

[0099] The test in Example 2 was repeated, with the difference that 90 people (whose symptoms of ankylosing spondylitis were not known before the test) were randomly selected for testing.

[0100] Prepare a test kit (100 person-times), which contains:

[0101]

[0102] 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 products of TAP1 and LMP2 were purified, they were sequenced with ABI 3730 DNA sequencer, and Polyphred software was used for sequence interpretation and SNP confirmation.

[0103] The results also confirmed that all individuals with "-- / AA / AA" tested had AS and that AS patients with G- / AG / AG had higher cases th...

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Abstract

The invention discloses a method for testing susceptibility of ankylosing spondylitis. The method comprises the following steps that: a TAPI gene or a transcript of an individual is detected and compared with a normal TAPI gene or a normal transcript; and simultaneously, an LMP2 gene or the transcript of the individual is detected and compared with a normal LMP2 gene or the normal transcript, wherein the existence of difference shows that the possibility of the individual with ankylosing spondylitis is higher than that of a normal group. The invention also discloses a corresponding detection 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 (single nucleotide polymorphism, SNP) of antigen peptide transporter 1 (Transporter of Antigen Peptides 1, TAP1) and low molecular weight polypeptide 2 (lowmolecular mass polypeptide-2, LMP2) and its relationship with tonic Association with 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 join...

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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