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Application of BMP2 and Noggin in jointly preparing ankylosing spondylitis diagnostic reagent kit

A technology for ankylosing spondylitis and a diagnostic kit, which is applied in the medical and health field, can solve the problems of difficult to reach the permissible range of the kit, difficult to detect serum, low serum level, etc., so as to reduce the lower limit of the permissible range of detection, improve the sensitivity, and improve the specificity. Effects of Sex and Sensitivity

Active Publication Date: 2016-07-27
沈慧勇
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, we found in experiments that although these kits can detect the levels of BMP2 and Noggin in cell culture supernatants, they are difficult to detect in serum. The reason is that the levels in serum are low and the tolerance range of the kits is difficult to reach this level

Method used

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  • Application of BMP2 and Noggin in jointly preparing ankylosing spondylitis diagnostic reagent kit
  • Application of BMP2 and Noggin in jointly preparing ankylosing spondylitis diagnostic reagent kit
  • Application of BMP2 and Noggin in jointly preparing ankylosing spondylitis diagnostic reagent kit

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Experimental program
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Effect test

Embodiment 1

[0040] Detection kit

[0041] (1) BMP2 detection kit, including: 96-well plate pre-coated with anti-human BMP2 antibody, 96-well plate film, BMP2 protein standard (1000pg / cart), standard diluent, biotin-labeled antibody, biological Corresponding index table of prime-labeled antibody diluent, horseradish peroxidase-labeled avidin, horseradish peroxidase-labeled avidin, TMB substrate chromogenic solution, stop solution, washing solution, and BMP2 detection values;

[0042] (2) Noggin detection kit, including: 96-well plate pre-coated with anti-human Noggin antibody, 96-well plate film, Noggin protein standard, standard diluent, biotin-labeled antibody, biotin-labeled antibody diluent , horseradish peroxidase-labeled avidin, horseradish peroxidase-labeled avidin diluent, TMB substrate chromogenic solution, stop solution, washing solution, Noggin detection value corresponding index table;

[0043] Preparation method of detection kit

[0044] (1) Antibody coating on 96-well p...

Embodiment 2

[0050] Application of detection kit in diagnosis of ankylosing spondylitis and prediction of severity of pathological osteogenesis

[0051] 1. The detection method of the detection kit of the present invention

[0052] (1) Extract the peripheral blood from AS patients, let it stand at room temperature for 1 hour, absorb the supernatant and centrifuge at 1000×g for 5 minutes, retain the supernatant serum, and discard the precipitate;

[0053] (2) Take a standard product from the kit, centrifuge it at 10000rpm for 1min, dissolve it with 1ml sample diluent, and dilute it 7 times (15.6-1000pg) by the doubling dilution method;

[0054] (3) Add 100ul of standard or test sample to each well, cover with film, and incubate at room temperature on a shaker for 2h;

[0055] (4) Discard the liquid, add 200 ul of washing solution to each well, and wash 3 times, each time for 1 min;

[0056] (5) Dissolve and dilute biotin-labeled antibody with biotin-labeled antibody diluent, add 100ul t...

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Abstract

The invention discloses application of BMP2 and Noggin in jointly preparing an ankylosing spondylitis diagnostic reagent kit.The invention further discloses application of BMP2 and Noggin in jointly preparing a reagent kit for predicting the severity of ankylosing spondylitis pathologic osteogenesis.The invention further discloses a BMP2 detection reagent kit and a Noggin detection reagent kit.By detecting the BMP2 level and the Noggin level at the same time, the specificity and sensitivity of diagnosis of ankylosing spondylitis can be improved, and the severity of ankylosing spondylitis pathologic osteogenesis can be predicted.By means of the BMP2 detection reagent kit and the Noggin detection reagent kit, the lower limit of the reagent kit detection allowable range is lowered, and the detection accuracy is improved.

Description

technical field [0001] The invention belongs to the medical and health field, and particularly relates to the application of BMP2 and Noggin in the preparation of an ankylosing spondylitis diagnostic kit. Background technique [0002] AS is a common autoimmune disease with a high prevalence rate of about 0.2-0.54% in my country (see: Rong, J.G. Jieruo, Spondyloarthritisinchina. CurrOpinRheumatol, 2013.25(4): p.460-7. ). AS tends to occur in young men and often involves the axial joints. The main clinical and pathological features are chronic progressive inflammation and pathological osteogenesis (see: Braun, J.J. Sieper, Ankylosingspondylitis. Lancet, 2007.369(9570): p.1379 -90.). However, the pathogenesis of AS is still unclear, which leads to the limitations and low effectiveness of current AS diagnosis and treatment methods. As the disease progresses, AS patients will gradually lose their ability to live and work, which brings heavy psychological pressure and economic b...

Claims

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Application Information

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IPC IPC(8): G01N33/68
CPCG01N33/6893G01N2800/10G01N2800/7095
Inventor 沈慧勇吴燕峰王鹏谢中瑜黄霖唐勇叶记超陈铿蔡兆鹏胡旭民李玉希
Owner 沈慧勇
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