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A biomarker for Behcet's disease detection and its application

A kind of use and technology of biological samples, applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of insufficient research specimens, incomplete diagnosis, unclear antibody positive rate and clinical application value, etc.

Active Publication Date: 2016-06-22
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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  • A biomarker for Behcet's disease detection and its application
  • A biomarker for Behcet's disease detection and its application
  • A biomarker for Behcet's disease detection and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Example 1 Screening of BD-related autoantibodies by high-density protein chips

[0046] All protein antigens on the high-density protein chip are N-terminal with GST tags for protein purification. First, the quality of the chip was verified by hybridization of rabbit anti-GST monoclonal antibody with the chip protein. Then, 40 BD patient sera, 15 autoimmune disease control sera and 20 healthy control sera were selected and hybridized with 75 high-density protein chips, and candidate BD-related autoantigens were identified through signal acquisition and data analysis.

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

[0048] The GenePix4000B chip scanner was used to scan the high-throughput protein chip to obtain a grayscale image in TIFF format. According to the operation manual of the GenePixPro6.0 software, according to the matrix parameters of the high-throughput chip loading point, the software automatically completes the signal pixel segmentation ...

Embodiment 2BD

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

[0064] Due to the differences in genetic backgrounds and environmental factors, the positive rates of autoantibodies produced by different patients vary greatly among patients with autoimmune diseases. In order to confirm whether the BD-related autoantibodies screened by high-density protein chip in small samples have clinical significance and application value, as well as the sensitivity and specificity of these antibodies in the diagnosis of BD patients, this study used high-density protein chips to screen BD-related autoantibodies. The related autoantibody target antigens were prepared into BD related autoantibody target antigen protein chips for clinical expansion sample verification.

[0065] Serum samples used for BD-related autoantibody target antigen protein chip for clinical large sample verification test included 130 BD patients, 103 autoimmune d...

Embodiment 3

[0073] Example 3 Immunoblotting verification of newly discovered BD autoantibodies

[0074] In order to verify whether the test results confirmed by the clinical large sample screening of BD autoantibody target antigen chip can be repeated in other detection methods, we used the most commonly used western blotting method in proteomics research methods to analyze the anti-CTDP1 antibody in the serum of BD patients. detection. The results of western blotting showed that the anti-CTDP1 antibody confirmed by the large sample screening of BD autoantibody target antigen chip could be well replicated in WB detection. There are autoantibodies that specifically react with CTDP1 in the serum of BD patients, and the response signal intensity is high. in the autoimmune disease control group and the normal control group. The results of immunoblotting detection are 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 biological marker for Behcet's disease detection and use thereof. Background technique [0002] Behcet's disease (BD), also known as Behcet syndrome (BS), is a common systemic autoimmune disease with unknown etiology, high incidence, recurrence, and coexistence of large and small vasculitis. . Three patients with a complex triad of oral and genital ulcers, hyphema, and uveitis were first described by Hulusi Behcet in 1937. Subsequent studies have shown that the disease is a systemic autoimmune disease with multi-system involvement and complex and variable clinical manifestations. Patients often have 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 mainly occurs in the Middle East and Mediterranean countries between 30 degre...

Claims

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Application Information

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Patent Type & Authority Patents(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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