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Hepatopathy detecting test paper by colloidal gold chromatography and preparation method thereof

A technology for detecting test strips and colloidal gold, applied in the field of medical immunity applications, can solve problems such as short detection time, and achieve the effects of short detection time, low production cost and low detection cost

Active Publication Date: 2011-05-18
SHANGHAI KEXIN BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are commercial ELISA kits on the market, but compared with gold standard chromatography test paper, there are still many differences. It is not limited by the site personnel. The detection time is as short as 5-10 minutes, and the interpretation of the results is simple.

Method used

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  • Hepatopathy detecting test paper by colloidal gold chromatography and preparation method thereof
  • Hepatopathy detecting test paper by colloidal gold chromatography and preparation method thereof
  • Hepatopathy detecting test paper by colloidal gold chromatography and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0063] Example 1 Antigen composition

[0064] The antigen protein used in this test paper is some components of the 2-oxo acid dehydrogenase complex [2-OADC] of the mitochondria for the target antigen of M2: pyruvate dehydrogenase (PDC-E2), 2-oxo acid Dehydrogenase complex E2 (BCOADC-E2), 2-oxoglutarate dehydrogenase complex E2 (OGDC-E2), through gene cloning technology to construct recombinant genes, and then use prokaryotic expression technology to successfully express all of human origin The three target antigen proteins of M2 were successfully connected to form an antigen protein triplet.

Embodiment 2

[0065] Example two antibody preparation

[0066] Antibody (a), antibody (c), and antibody (b) are prepared by the following methods to realize the preparation of colloidal gold chromatography test paper for liver disease detection. Among them, anti-human IgG, antibody (c) and antibody (b) can generally be produced by multiple subcutaneous (sc) or intraperitoneal (ip) injections of purified immunogen and adjuvants on animals.

[0067] By mixing 0.05mg~1mg immune preparations (respectively for goats or mice) with 3 times the volume of Freund's complete adjuvant, the solution is injected into multiple sites in the skin. One month later, 1 / 5 to 1 / 10 of the animal The original amount of Freund's complete adjuvant mixture of human IgG is boosted by subcutaneous injection of multiple sites. After 7 to 14 days, the animals were bled to determine the serum anti-human IgG titer. Boost animals until the titer reaches a plateau. The methods of producing polyclonal antibodies are described i...

Embodiment 3

[0069] Example Three sheep anti-human IgG labeling

[0070] Labeling of goat anti-human IgG: adjust the pH of colloidal gold to 8.0~9.0 with 0.1M potassium carbonate, slowly add 8~12μg goat anti-human IgG per ml of colloidal gold solution, stir for 10~30min, then add BSA to a final concentration of 0.5~1 %, stir for 10-30min, centrifuge, discard the supernatant, wash the precipitate with the washing solution for 2 to 3 times, and resuspend the precipitate with one-tenth of the initial volume of the preservation solution at the end, and store it at 4°C for use.

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Abstract

The invention discloses hepatopathy detecting test paper by colloidal gold chromatography and a preparation method thereof. The hepatopathy detecting test paper by colloidal gold chromatography is a film tape formed by sequentially and mutually overlapping and pasting a pyroxylin coating film, a bonding pad, a sample pad and a water-absorption pad, wherein the pyroxylin film is provided with a detecting line and a quality control line, the detecting line coats M2 type mitochondrial antigenic protein, the quality control line coats an antibody (c), and the bonding pad coats a colloidal gold labeled antibody (a) and a colloidal gold labeled antibody (b). In the test paper tape provided by the invention, the M2 type mitochondrial antigenic protein is introduced, and process optimization is carried out on the bonding pad and the sample pad, thus realizing detection performance with high sensitivity, high specificity and high accuracy on the M2 type anti-mitochondrial antibody.

Description

Technical field [0001] The invention belongs to the field of medical immunity application, and specifically relates to a test paper for detecting liver diseases by using colloidal gold immunochromatography technology and a preparation method thereof. Background technique [0002] Primary biliary cirrhosis (Primary biliary cirrhosis, PBC) is an organ-specific autoimmune liver disease that mainly destroys the small and medium-sized bile ducts in the liver. It is manifested as inflammation of the portal vein area leading to fibrosis, cirrhosis and even function. Sexual failure. In patients with hepatitis and liver cirrhosis, various pathogen detection indicators are negative, and it is necessary to consider primary biliary cirrhosis or autoimmune hepatitis. Early use of drug therapy can control the progression of the disease. The key to the treatment of PBC is early treatment. The premise of early treatment is early diagnosis, and early diagnosis has become the focus of everyone's ...

Claims

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Application Information

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IPC IPC(8): G01N33/576G01N33/577G01N33/532
CPCG01N33/558G01N33/54386G01N33/54388
Inventor 张玥韩永俊高成秀杨超文孙宏彬孙潇张宇婷钱杰
Owner SHANGHAI KEXIN BIOTECH
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