Kit for detecting LKM (anti-liver-kidney microsomal antibody)-1 relative to autoimmune liver diseases, and detection method with kit

A technology of autoimmunity and reagent kits, applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve problems such as differences and difficulties in development

Inactive Publication Date: 2013-03-06
AUTOBIO DIAGNOSTICS CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Clinically, indirect immunofluorescence (IIF) is commonly used to detect anti-LKM-1 antibodies in patients. The experimental matrix is ​​usually frozen sections of liver, stomach, and kidney tissues of different primates, which contain a complete antigen spectrum, but for indirect Immunofluorescence analysis staining identification has a certain degree of subjectivity, so the interpretation of the results may vary at different time points, between different observers, and between old and new tissue sections, and requires the assistance of fluorescent microscopes and other instruments. General small and medium hospitals are difficult to carry out

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] The kit (plate type) for detecting autoimmune liver disease-related antibodies and anti-LKM-1 antibodies of the present invention includes:

[0022] 1. Preparation of target antigen-carrier conjugate: Add 100 μl of target antigen (natural antigen, recombinant antigen, synthetic peptide, HepG2 cell, Hep2 cell, Hela cell or cell lysed antigen) not higher than 10ug / ml to a 96-well plate In the medium, coat at 4°C for 16-24h or at 37°C for 2h, then wash twice with the prepared (PBST) washing solution;

[0023] 2. Blocking: Use 50-200 μl of 0.01-0.1M phosphate buffer containing 0.05-0.5% Tween20, 1-5% BSA, 1-5% Casein, 1-5% peptone, 5-10% sucrose, Block at 4°C for 16-24h or at 37°C for 2h, then shake off excess buffer, place in a dry environment to dry, and seal at 2-8°C for storage;

[0024] 3. Preparation of calibrator, positive control substance and negative control substance: take a clinically determined high-value sample, dilute it with the calibrator diluent (PBS+3%BS...

Embodiment 2

[0039] The kit (magnetic particle) for detecting autoimmune liver disease-related antibody anti-LKM-1 antibody described in the present invention includes:

[0040] 1. Preparation of target antigen-carrier conjugates: Take 10mg of magnetic particles, wash them with phosphate buffer for 5 times, add no less than 0.15ml of EDC or (and) NHS or glutaraldehyde (and) activator for 2 hours, wash 3 times, then add 0.1mg of target antigen, shake at room temperature for 2h;

[0041] 2. Blocking: Use 1ml of 0.01-0.1M phosphate buffer containing 0.05-0.5% Tween20, 1-5% BSA, 1-5% Casein, 1-5% peptone, 5-10% sucrose, shake at room temperature Block for 10 minutes, block four times in a row, then add 2ml of blocking solution and store at 2-8°C;

[0042] 3. Preparation of calibrator: Take a high-value sample with a clinically determined value, dilute it with a calibrator diluent at a certain ratio, and take the point with better linearity as the calibrator;

[0043] 4. Preparation of the en...

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Abstract

The invention discloses a kit for detecting LKM (anti-liver-kidney microsomal antibody)-1 relative to autoimmune liver diseases, and a quantitative and quantitative detection method for the LC-1 antibody with the kit. The detection method comprises the following steps of: combining a sample with IgG or / and anti-human IgM or / and anti-human IgA, which is / are connected with marker(s), and specific antigen corresponding to the detected antibody; adding a substrate of a luminescence system or a developing system or a light exciting system into the sample, and then reading a signal value of the sample to be detected; and calculating a quantitative and quantitative result or reading a quantitative and quantitative result of the sample by naked eyes or instruments according to immunochromatography or immuno-infiltration, thereby accurately diagnosing the type and severity of the autoimmune liver diseases to guide a doctor to dispense medicines. The quantitative and quantitative detection method has the advantages of high sensitivity, strong specificity and high accuracy, and is applicable to interpretation by semi-automatic or full-automatic detection systems or naked eyes.

Description

technical field [0001] The invention relates to the field of biological detection, in particular to a kit for detecting autoimmune liver disease-related antibody anti-LKM-1 antibody, and also relates to a method for using the kit to detect anti-LKM-1 antibody. Background technique [0002] Autoimmune liver diseases (autoimmune liver diseases) mainly include three diseases that are closely related to autoimmunity and mainly focus on liver and gallbladder damage: autoimmune hepatitis (AIH), primary biliary cirrhosis (primary biliary cirrhosis) , PBC) and primary sclerosing cholangitis (primary sclerosing cholangitis, PSC). AIH is a liver disease with circulating autoantibodies and hyperimmunoglobulinemia, unknown etiology, and chronic inflammatory necrosis. PBC is an autoimmune-mediated intrahepatic bile duct injury, followed by liver fibrosis, and eventually leads to hepatic fibrosis. PSC is a chronic syndrome of unknown etiology characterized by chronic cholestasis caused b...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/576G01N21/76
Inventor 吴学炜张学东于林何丽鲁清月杨皓宇付光宇马建军刘功成项立红渠海李桂林
Owner AUTOBIO DIAGNOSTICS CO LTD
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