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Novel test method for rheumatoid arthritis and kit for rheumatoid arthritis test

A rheumatoid, inspection method technology, applied in biochemical equipment and methods, microbial determination/inspection, biological testing, etc.

Active Publication Date: 2013-03-13
KAYTEE BIO
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, in the reports so far, the sensitivity of rheumatoid factor is 75-80% and the specificity is 50-70%, and the sensitivity of anti-CCP antibody is 50-75% and the specificity is 85-95%. Meet needs (see non-patent literature 2,3)

Method used

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  • Novel test method for rheumatoid arthritis and kit for rheumatoid arthritis test
  • Novel test method for rheumatoid arthritis and kit for rheumatoid arthritis test

Examples

Experimental program
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Embodiment 1

[0045] The blood of each subject was collected into an EDTA tube, and centrifuged at 2500 rpm for 10 minutes at room temperature to obtain plasma.

[0046] The amount of talin in the above plasma was measured by sandwich ELISA method.

[0047] First, the H-18 antibody (Santa Cruz Biotechnology Co., Ltd.) that recognizes the N-terminus of talin was diluted to 1 μg / mL with phosphate buffered solution (PBS), and then added to a 96-well microwell plate in an amount of 100 μL / well. Incubate overnight at 4°C, and then wash 3 times with 200 μL / well of washing solution. Next, 100 μL / well of plasma from each subject was added to a 96-well microplate, incubated at 25° C. for 1 hour, and then washed 3 times with 200 μL / well of washing solution. Next, the H-300 antibody (Santa Cruz Biotechnology Co., Ltd.) that recognizes the N-terminus of talin as the primary antibody was diluted with PBS to make it 2 μg / mL, and then added to a 96-well microplate in an amount of 100 μL / well Incubate at...

Embodiment 2

[0054] The amount of talin in plasma was measured by the sandwich ELISA method in the same manner as in Example 1, except that the M54246M antibody (Biodesign) recognizing the C-terminus of talin was used as the primary antibody.

[0055] The ROC curve of embodiment 2 is as figure 2 shown. The results of the ROC analysis, figure 2 The area under the ROC curve (AUC) was 0.819. In addition, the positive and negative results when OD=0.05 was used as the cutoff value are shown in Table 2 below.

[0056] [Table 2]

[0057]

[0058] From these results, the sensitivity of rheumatoid arthritis diagnosis by the sandwich ELISA method using the H-18 antibody and the M54246M antibody was 15 / 17×100=88.2%, and the specificity was 11 / 14×100=78.6%. .

Embodiment 3

[0067] The therapeutic effects of rheumatoid arthritis therapeutic agents on 5 RA patients were confirmed by measuring the amount of talin. As in Example 1, the amount of talin was measured by measuring the OD value by the sandwich ELISA method using the H-18 antibody and the H-300 antibody. In addition, the CRP level and the MMP-3 level were also measured by the same method as the usual clinical examination. Furthermore, the DAS (Disease Activity Score) 28 score recommended by the European Union for the Prevention and Treatment of Rheumatism (EULAR) was also calculated. In terms of DAS28 scores, 5.1 or more is judged as high disease activity, 3.2 or more and less than 5.1 is judged as moderate disease activity, and less than 3.2 is judged as low disease activity. The results are shown in Table 4 below.

[0068] [Table 4]

[0069]

[0070] Case 1 is an example of a good responder (DAS28: 5.43→2.62) when EULAR response criteria were applied by using ADA (adalimumab) in co...

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Abstract

Provided are: a novel test method for rheumatoid arthritis; and a kit for rheumatoid arthritis test, which is used in the novel test method for rheumatoid arthritis. A test method for rheumatoid arthritis according to the present invention is characterized by comprising a step for measuring the amount of talin in the plasma or serum of an animal subject. This measurement is carried out, for example, by an immunological method using an antibody which binds to talin. A kit for rheumatoid arthritis test according to the present invention is used for such a test method and contains, for example, a solid-phase carrier to which an antibody that binds to talin is affixed.

Description

technical field [0001] The present invention relates to a method for examining rheumatoid arthritis, and a kit for examining rheumatoid arthritis used in the method for examining rheumatoid arthritis. Background technique [0002] Rheumatoid arthritis (Rheumatoid Arthritis: RA) is a chronic inflammatory disease in which the synovial tissue of the joint is the main lesion site, and the prevalence rate accounts for about 1% of the population. Rheumatoid arthritis causes synovitis in its initial stage, then invades cartilage / bone, and when it progresses, the joint is destroyed and deformed. In addition, there are many cases in which symptoms progress, such as repeated remission / relapse of arthritis, cases of recovery, cases of rapid deterioration, and the like. [0003] Diagnosis of rheumatoid arthritis is mainly based on symptoms, but in recent years, a diagnostic method using autoantibodies contained in patient serum as markers has attracted attention. As such autoantibodie...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/53G01N33/564
CPCG01N33/564G01N2800/102C12Q1/6886G01N2333/4703
Inventor 津坂宪政
Owner KAYTEE BIO
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