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ACPA-negative RA diagnosis marker and application thereof

A negative and useful technology, applied in the field of RA diagnostic markers, which can solve problems such as molecular characteristics of affected joints, differences in remission rate and response rate, etc.

Active Publication Date: 2017-07-14
北京埃克帕生物医学科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, ACPA-positive and ACPA-negative RA patients differ markedly in the genetic and environmental determinants of the disease, the molecular characteristics of the affected joints, the rate of remission and, most importantly, the rate of response to treatment

Method used

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  • ACPA-negative RA diagnosis marker and application thereof
  • ACPA-negative RA diagnosis marker and application thereof
  • ACPA-negative RA diagnosis marker and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0054] Example 1 High-density protein chip screening candidate RA autoantigens

[0055] The high-density protein chip and the expression recombinant plasmid of Saccharomyces cerevisiae containing the target gene sequence were provided by the laboratory of Professor Zhu Heng of Johns Hopkins University in the United States. Each high-density protein chip contains 48 microarrays (blocks), and each microarray contains 992 probe points, arranged in a 32*31 array, and each protein probe on the chip has 2 parallel points. The protein chip contains 21827 non-redundant recombinant human proteins. The recombinant protein comes from the full-length open reading frame (open reading frame, ORF) of the corresponding gene expressed by the Saccharomyces cerevisiae host, and its N-terminus has a glutathione S-transferase (glutathione S-transferase, GST) tag.

[0056] Use the mouse anti-GST monoclonal antibody to hybridize with the chip to verify the quality of the chip. When the correlation ...

Embodiment 2

[0058] Example 2 Hybridization of high-density protein chips with RA and control serum

[0059] Select 60 RA sera and 60 control sera (10 Behcet's disease sera, 10 Takayasu arteritis sera, 10 SLE sera and 30 healthy human sera) to hybridize with 120 chips, and identify candidates through signal acquisition and data analysis RA autoantigens. Anti-human IgG antibodies labeled with PE-Cy5 were used to detect the reaction of autoantibodies in serum with corresponding autoantigen probes. image 3 Shown are the representative local image results after the high-density protein chip was reacted with serum, and the different protein antigen probes are in the box. A, C, E, and G show schematic diagrams of hybridization of four RA sera to the chip. B, D, F, H show the schematic diagrams of hybridization of four control sera (including healthy control and disease control) with the chip. Panel I is a schematic diagram of RA that is effective in treatment, and panel J is a schematic diag...

Embodiment 3RA

[0075] Example 3 Construction of RA autoantigen protein chip and verification of serum screening

[0076] A total of 46 candidate RA autoantigens were screened by analyzing the hybridization results of high-density protein chip and small sample serum. In order to verify the specificity and sensitivity of these autoantigens, the present invention prepared a RA autoantigen protein chip with low probe density. Table 2 is the microarray layout of each probe on the RA autoantigen protein chip. The probes on the chip included 46 candidate RA autoantigens screened by the large chip and 5 control IGHG1 probes.

[0077] The microarray layout of each probe on the table 2RA autoantigen protein chip

[0078] AK2 IGHG1 ND ATP13A5 ND TBC1D19 RAB35 UBXN10 RAB3D APH1A TNFAIP1 HDAC4 ARL2BP RAI14 RRN3 POLR3B ERH NDRG1 BLANK BLANK BLANK BLANK BLANK GARS SUGT1 IGHG1 NOL3 ZSCAN20 LSP1 RGCC EMPTY PAGE5 FGF12 ...

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Abstract

The invention provides applications of deoxyhypusine dioxygenase, namely, DOHH, or a fragment thereof in preparation of a reagent for diagnosis of anti-citrullinated protein antibody negative (ACPA-negative) rheumatoid arthritis (RA). In the method, a high-density protein chip is hybridized with RA serum to screen 35 proteins, which are candidate auto-antigens of the ACPA-negative RA. It is identified that four protein antigens, DOHH, DUSP11, PTX3 and PAGE5, have high sensitivity and specificity in ACPA-negative serum of the RA, wherein the four protein antigens are all novel auto-antigens. The method shows that the DOHH, DUSP11, PTX3 and PAGE5 can be used as diagnosis markers for the ACPA-negative RA.

Description

technical field [0001] The invention belongs to the field of biological detection, and in particular relates to an ACPA-negative RA diagnostic marker and application thereof. Background technique [0002] Rheumatoid arthritis (Rheumatoid arthritis, RA) is a chronic autoimmune disease, which mainly causes inflammation in the local synovium of multiple small joints throughout the body, and then causes local bone destruction in the joints as a group of clinical manifestations disease. Rheumatoid arthritis affects approximately 0.5%-1% of the population in developing countries. Overall, RA is more common in women than in men, and more common in the elderly than in the young. The clinical presentation of rheumatoid arthritis varies widely, ranging from a self-limited disease with mild symptoms to rapidly progressive inflammation with joint destruction and severe physical disability. Because of the variability in disease manifestations, classification criteria have been develop...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/564G01N33/68
CPCG01N33/564G01N33/6893G01N2800/102G01N33/573G01N2333/90241
Inventor 张烜莫文秀李永哲胡朝军刘国振武丽君
Owner 北京埃克帕生物医学科技有限公司
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