Biomarker for Behcet disease (BD) detection and use thereof

A technology of purpose and subunit, applied in the field of biological detection, can solve the problems of insufficient research specimens, imperfect diagnosis, inability to determine the positive rate of antibodies and clinical application value, etc.

Active Publication Date: 2015-07-15
PEKING UNION MEDICAL COLLEGE HOSPITAL CHINESE ACAD OF MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In 2014, Morishima used the protein chip technology containing 9,000 human gene-encoded proteins to conduct a screening study on the serum of BD patients. The results of the study showed that there were anti-tight junction protein-1 antibodies in BD patients. Due to the insufficient amount of research samples involved in this study, The positive rate and clinical application value of this antibody in BD patients are still unclear
Current studies have shown that if relying on clinical symptoms, imaging examinations, and serum autoantibody examinations, only some BD patients can be detected and diagnosed correctly in time, and the diagnosis of BD is still not perfect.

Method used

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  • Biomarker for Behcet disease (BD) detection and use thereof
  • Biomarker for Behcet disease (BD) detection and use thereof
  • Biomarker for Behcet disease (BD) detection and use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Example 1 High-density protein chip screening for BD-related autoantibodies

[0046] The N-terminal of all protein antigens on the high-density protein chip has a GST tag for protein purification. First, use rabbit anti-GST monoclonal antibody to hybridize with chip protein to verify chip quality. Then, 40 BD patient sera, 15 autoimmune disease control sera, and 20 healthy control sera were selected for hybridization with 75 high-density protein chip chips, and candidate BD-related autoantigens were identified through signal collection and data analysis.

[0047] 1. Extraction of high-density protein chip scanning data

[0048] Use the GenePix 4000 B chip scanner to scan the high-throughput protein chip, and obtain the grayscale image in TIFF format. According to the operation manual of the GenePix Pro 6.0 software, and according to the high-throughput chip loading matrix parameters, the software automatically completes the signal of each protein point. Pixel segmenta...

Embodiment 2

[0063] Example 2 Construction of BD-related autoantibody target antigen protein chip and verification of serum screening

[0064] Due to differences in genetic background and environmental factors in patients with autoimmune diseases, the positive rate of autoantibodies produced by different patients varies greatly. In order to confirm whether the BD-related autoantibodies screened out by high-density protein chips in a small sample have clinical significance and application value, and the sensitivity and specificity of these antibodies to the diagnosis of BD patients, this study used the BD-related autoantibodies screened out by high-density protein chips Related autoantibody target antigens were prepared into BD-related autoantibody target antigen protein chips for clinical expansion sample verification.

[0065] The serum samples used for BD-related autoantibody target antigen protein chips for clinical large-scale verification and detection included 130 BD patients, 103 au...

Embodiment 3

[0073] Example 3 Western blot verification of newly discovered BD autoantibodies

[0074] In order to verify whether the test results confirmed by BD autoantibody target antigen microarray clinical large sample screening can be replicated by other detection methods, we used the most commonly used method in proteomics research methods to detect anti-CTDP1 antibodies in serum of BD patients. detection. The results of western blotting showed that the anti-CTDP1 antibody confirmed by the large-scale screening of BD autoantibody target antigen chip can be well repeated in WB detection, and there are autoantibodies that specifically react with CTDP1 in the serum of BD patients, and the reaction signal intensity is high In autoimmune disease control group and normal control group. The results of western blot detection were as Figure 4 .

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Abstract

The invention provides a use of RNA polymerase II subunit A C-terminal domain phosphatase (CTDP1) or a fragment thereof in preparation of a reagent used for diagnosing Behcet disease (BD). The positive rate of the CTDP1 or the fragment thereof in a BD patient is remarkably higher than that of normal control, a Western blot verification result displays that an anti-CTDP1 antibody can be detected in the serum of the BD patient, which proves that the CTDP1 or the fragment thereof can be used for a detection marker of BD.

Description

technical field [0001] The invention belongs to the field of biological detection, and in particular relates to a biomarker for Behcet's disease detection and its application. Background technique [0002] Behcet's disease (BD), also known as Behcet syndrome (BS), is a common systemic autoimmune disease with unknown etiology, high incidence, relapse, and coexistence of large and small vasculitis . In 1937 Hulusi Behcet first described three patients with a complex triad of oral and genital ulcers, hypopyon, and uveitis. Subsequent studies have shown that the disease is a systemic autoimmune disease with multi-system involvement and complex and variable clinical manifestations. Patients often present with recurrent oral and genital ulcers, arthritis, uveitis, skin lesions, thrombophlebitis, and gastrointestinal and central nervous system involvement. BD can be found all over the world, but it mainly occurs in the Middle East and Mediterranean countries located between 30 d...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/558G01N33/532
CPCG01N33/532G01N33/558
Inventor 李永哲胡朝军朱衡宋光吴子燕陈思张奉春
Owner PEKING UNION MEDICAL COLLEGE HOSPITAL CHINESE ACAD OF MEDICAL SCI
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