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Detection method and reagent kit of anti-keratin antibody

An anti-keratin, detection method technology, applied in biological testing, material inspection products, fluorescence/phosphorescence, etc., can solve the problems of weak fluorescence intensity, missed detection, etc., achieve increased fluorescence intensity, convenient use, improved resolution and The effect of sensitivity

Inactive Publication Date: 2008-12-10
TIANJIN BAODI HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] However, only IgG anti-keratin antibodies can be detected by using the anti-keratin antibody kit of Oumeng Company in Germany, and IgG anti-keratin antibodies only account for about 85% of all antibodies, so there are still about 15% of IgA and IgM Patients with type anti-keratin antibodies will be missed, and this method has weak fluorescence intensity and strong non-specific fluorescence, which may cause missed detection

Method used

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  • Detection method and reagent kit of anti-keratin antibody
  • Detection method and reagent kit of anti-keratin antibody
  • Detection method and reagent kit of anti-keratin antibody

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] The detection method of embodiment 1 anti-keratin antibody

[0030] During detection, add 25 μl of 1:10 diluted serum to be tested in the reaction area of ​​the biological sheet, incubate in a wet box at 37°C for 30 minutes, rinse with PBS, spin dry, add 25 μl of complement, incubate in a wet box at 37°C for 30 minutes, rinse with PBS, Shake dry, then add fluorescein-labeled anti-C 3 c antibody 25 μl, incubated at 37°C for 30 minutes, rinsed with PBS, dried, and mounted with buffered glycerol. Mounting is not required for direct detection. The slides were observed under a fluorescent microscope, and the typical regular linear or lamellar fluorescence in the stratum corneum was positive.

[0031] In order to better understand the present invention, the positive effect of the present invention aspect detection anti-keratin antibody is further illustrated by the result that two kinds of detection methods draw during clinical detection below:

[0032] The selected cases ...

Embodiment 2

[0042] Example 2 Kit

[0043] 1. Box body

[0044] The kit of the present invention is for one-time use. The box body 1 is in the shape of a hexahedron made of cardboard and consists of a box bottom 5 and a box cover 2 . The bottom of the box is connected with the lid as a whole. Bottom of the box 5 is provided with a bottle holder 3 at the long side where the bottom of the box 5 links to each other with the lid 2, and the bottle holder is provided with 7 holes 4 for placing bottles 10 that various reagents are housed. In the middle of the bottom of the box 5 there is a baffle 7 extending from the bottle holder 3 to the other long side, dividing the bottom of the box 5 into two parts, one of which is placed with a buffer and a cover, and the other part is placed with a carrier sheet with a packaging bag 6 13 and instructions (not shown). The length, width and height of the box body match the shape of the contained items.

[0045] 2. Items equipped with the kit

[0046] 1...

Embodiment 3

[0056] Embodiment 3 kit experimental operation steps

[0057] Preparation: Take the slides out of the kit and let it warm to room temperature.

[0058] Dilution: first dilute 50ml of concentrated phosphate buffer solution with distilled water to 1L, the phosphate content of the diluent is 10.2g, pH=7.2. Add 2ml Tween 20 into the diluted phosphate buffered saline (PBS-Tween 20), and stir well. Dilute the serum to be tested with PBS-Tween 20 at a ratio of 1:10 (positive and negative control serum do not need to be diluted), and mix well before use. Reconstitute 100 μg of powdered complement with 50 μl of distilled water into a complement application solution.

[0059] Loading: Add 25ul of diluted serum dropwise to the reaction area of ​​the slide to avoid air bubbles. For the first incubation: place the slide with the bioflake facing up, cover the reaction area with a cover slip, and the reaction starts immediately. Make sure the specimen is in contact with the biochip and i...

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Abstract

The invention discloses an anti-keratin antibody detection method and an anti-keratin antibody regent box, and belongs to the method for detecting the characteristics of blood in body. The method provided by the invention can be operated as follows: the esophagus of a bandicoot can be made into a biological thin slice which can be taken as an antigen and coated in the reaction region of a piece of slide glass; blood serum to-be-detected is added in the reaction region and then added with complement after the incubation, rinsing and drying in a spanning way, then added with the fluorescent markers of anti-complement antibodies after the incubation, rinsing and drying in a spanning way, and then observed under a fluorescence microscope after the incubation, rinsing and drying in a spanning way; if the horny layer appears typical and regular line-shaped or lamellar fluorescence, the detection result is positive. The regent box provided by the utility model is filled with regents, cover glass and slide glass with biological thin slices, such as negative control serum, positive control serum, condensed phosphate buffer solution and the application solution of the fluorescent markers of complement and anti-complement antibodies. The method provided by the invention is better than the prior method in indicators such as sensitivity, specificity, negative predictive value and positive predicative value, therefore, the method has higher diagnosis value in diagnosing rheumatoid diseases.

Description

technical field [0001] The invention relates to a method for measuring blood characteristics in vivo, in particular to a detection method and kit for anti-keratin antibodies Background technique [0002] Rheumatoid arthritis (RA) is a chronic systemic autoimmune disease characterized by joint synovitis. Synovitis persists and recurs, which can lead to the destruction of cartilage and bone in the joint, joint dysfunction, and even disability. Vasculitis lesions involve various organs of the whole body, so the disease is also called rheumatoid disease. At present, the clinical diagnosis of RA mainly relies on the rheumatoid diagnostic criteria revised by the American Rheumatology Association (ARA) in 1987, including: clinical manifestations, X-ray changes, and rheumatoid factor (RF) detection, and patients who meet the ARA criteria are basically It is the advanced or late stage of RA, but early detection, early diagnosis and early active and effective treatment can greatly r...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/53G01N21/64
Inventor 王高生李立和
Owner TIANJIN BAODI HOSPITAL
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