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Kit for joint or single detection of autoimmune liver disease related antibody and detection method of kit

An autoimmune, kit-based technology that can be used in measurement devices, instruments, scientific instruments, etc., to solve problems such as poor prognosis and increased liver cirrhosis

Inactive Publication Date: 2012-10-03
AUTOBIO DIAGNOSTICS CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The probability of liver cirrhosis in PBC patients positive for this antibody is significantly increased, the serum bilirubin is elevated, the disease progresses rapidly, and the prognosis is poor

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] The kit (plate type) for joint detection of autoimmune liver disease-related antibodies ANA and ASMA of the present invention includes:

[0034] 1. Preparation of target antigen-carrier conjugates: one or more target antigens (natural antigens, recombinant antigens, synthetic peptides) that can specifically bind to ANA and ASMA antibodies, and the concentration is not higher than 10ug / ml , HepG2 cells, Hep2 cells, Hela cells or cell lysate antigens) each 100μl was added to a 96-well plate, coated at 4°C for 16-24h or 37°C for 2h, and then washed with the prepared (PBST) washing solution for two all over;

[0035] 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;

[0036] 3. Preparation of calibrator, positive control substance and negative cont...

Embodiment 2

[0051] The kit (magnetic particle) for joint detection of autoimmune liver disease-related antibodies ANA and ASMA of the present invention includes:

[0052] 1. Preparation of target antigen-carrier conjugates: Take 10 mg of magnetic particles, wash them with phosphate buffer for 5 times, add no less than 0.15 ml of EDC or (and) NHS or glutaraldehyde (and) activator for 2 hours, Wash 3 times, then add 0.1mg of ANA antigen and ASMA antigen respectively, shake at room temperature for 2h;

[0053] 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;

[0054] 3. Preparation of calibrator, positive control substance and negative control substance: take clinically high-value samples with ANA and ASMA values, and dilute them with calibrator diluent (PBS+3%BSA) at no less than ...

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Abstract

The invention discloses a kit for joint or single detection of an autoimmune liver disease related antibody, and a method for quantitative and qualitative detection of the autoimmune liver disease related antibody by using the kit. A sample is combined with a specific antigen which is connected with IgG or / and anti-human IgM or / and anti-human IgA of a label and corresponds to a detection antibody, a substrate of a luminescent system or a color development system or an exciting light system is added, then the signal value of the sample to be tested is read, and a quantitative or qualitative result is calculated or the quantitative or qualitative result is read through an immuno-chromatographic or an immuno-percolation method naked eye or an instrument, so that the type and the severity of an autoimmune liver disease are accurately diagnosed, medication performed by doctors is guided, and the method is high in sensitivity, strong in specificity and high in accuracy, and is suitable for a semi-automatic or full-automatic detection system or naked eye interpretation.

Description

technical field [0001] The invention relates to the field of biological detection, in particular to a kit for combined or independent detection of autoimmune liver disease-related antibodies, and also relates to a method for using the kit to detect related antibodies. 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 ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/576G01N33/543
Inventor 张学东付光宇杨皓宇于林何丽吴学炜马建军刘功成项立红渠海李桂林
Owner AUTOBIO DIAGNOSTICS CO LTD
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