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Self-immunity hepatitis detection protein chip and kit thereof

An autoimmune, protein chip technology, applied in the field of autoimmune hepatitis detection protein chips and kits, can solve the problems of lagging detection indicators, lack of integration, and single detection function.

Inactive Publication Date: 2010-06-23
上海裕隆生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Solve the shortcomings of single detection function, lagging detection indicators, and no integration of autoimmunity detection products on the market

Method used

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  • Self-immunity hepatitis detection protein chip and kit thereof
  • Self-immunity hepatitis detection protein chip and kit thereof
  • Self-immunity hepatitis detection protein chip and kit thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Preparation of a protein chip for detecting autoimmune hepatitis

[0036] The main steps are: (1) Prepare the matrix nitrocellulose membrane; (2) Design the chip, determine the arrangement of the lattice and the spotting position; (3) Draw the antibody from the 384-well plate and spray it on the nitrocellulose (4) assemble a nitrocellulose membrane, and carry out sealing, drying, packaging, and preservation.

[0037] The preparation method of the above-mentioned protein chip for detecting multiple AIH marker antibodies specifically includes the following steps:

[0038] 1. Determination of autoimmune hepatitis marker detection antigen antibody:

[0039] For the preparation of autoimmune hepatitis detection protein chip detection index antigens, mainly by extraction or recombination, the source of each index is shown in the table below:

[0040] antigen

source

antigen

source

dsDNA

plasmid

RNP (proteins A, C, 68kDa)

recombinant ex...

Embodiment 2

[0049] Kit preparation

[0050] A protein chip kit for detecting autoimmune hepatitis, which is composed of a protein chip for detecting autoimmune hepatitis, a standard product, a quality control product, a sample diluent, an enzyme-labeled working solution, a concentrated washing solution, and a detection solution; each part is bottled, Fix it in the box with a paper template (such as figure 2 ), not limited to the placement method, it is used together with the test. Here's how the parts are prepared:

[0051] 1. The autoimmune hepatitis detection protein chip is a 6×8 well plate, with nitrocellulose membrane as the base, and the assembly method is shown in ZL032298900. After the completion of the appearance, see figure 1 .

[0052] 2. The standard product provides 6 bottles, each bottle is 0.25ml.

[0053] With 3% BSA and 0.05% preservatives (Proclin TM 300) phosphate buffer (pH 7.4) configuration, where Anti-dsDNA, Anti-RNP, Anti-Sm, Anti-SS-A / Ro, Anti-SS-B / La, Anti...

Embodiment 3

[0072] Application and detection of protein chips and kits

[0073] The application of the protein chip for detecting autoimmune hepatitis of the present invention mainly uses the antigen-antibody-enzyme-labeled secondary antibody method to detect autoimmune hepatitis marker antibodies in human blood. The method is to add human serum to the surface of the chip, and the antibodies in the sample are respectively combined with the corresponding antigens immobilized on the chip; after drying, add the enzyme-labeled secondary antibody to react, and wash away the unbound secondary antibody; after adding the detection solution, 1 Minutes, scan and collect the signal, the signal intensity of the sample point is proportional to the concentration of the antibody in the serum.

[0074] In the application of the above-mentioned protein chip kit for detecting autoimmune hepatitis, the specific methods for its antigen-antibody reaction and detection are as follows:

[0075] (1) The chips s...

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Abstract

The invention discloses a self-immunity hepatitis detection protein chip and a kit thereof. The chip and the kit can be used for simultaneously detecting 12 immunity protein markers: ANA (dsDNA, RNP, Sm, SS-A / Ro, SS-B / La, Sc1-70, Jo-1), SMA, LKM-1, SLA / LP, F-actin and AMA. The invention overcomes the shortcomings of the current products that the detection index is single and incomplete. The invention provides a detection method which has a complete diction result and a simpler process.

Description

technical field [0001] The invention relates to a protein chip disease detection product, in particular to a protein chip for autoimmune hepatitis detection and a kit thereof. Background technique [0002] Autoimmune liver diseases mainly include primary biliary cirrhosis (PBC), autoimmune hepatitis (AIH), primary sclerosing cholangitis (PSC), and the three Overlap syndrome between any two of the disorders. Although they are all autoimmune liver diseases, their etiology, clinical manifestations, and treatment are different, and early diagnosis and early treatment are the key to controlling disease progression, so timely diagnosis and differential diagnosis have become the primary measures of clinical treatment. Laboratory tests, especially the determination of immune indicators, play an important role in clinical diagnosis and differential diagnosis. [0003] At present, the AIH scoring system developed by the International Autoimmune Hepatitis Group (IAIHG) is generally u...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/576
Inventor 穆海东汪宁梅穆宇豪刘纲
Owner 上海裕隆生物科技有限公司
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