Encapsulation method of bacterial lipopolysaccharide, reagent kit and method of detecting specific lipopolysaccharide

A bacterial lipopolysaccharide and kit technology, which is applied in the field of bacterial lipopolysaccharide coating, can solve problems such as difficulty in embedding LPS antigens and affecting the stability of ELISA test results.

Inactive Publication Date: 2002-10-16
SOUTH CHINA SEA INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] The purpose of the present invention is to overcome the shortcoming that indirectly affects the stability of its ELISA detection results because of the difficulty in embedding the LPS anti

Method used

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Examples

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Embodiment Construction

[0051] The following combined examples are to further illustrate the present invention, but should not be regarded as limiting the present invention.

[0052] The coating method of bacterial LPS is

[0053] (a) 0.8 μg LPS was dissolved in 100 μl pH9.6 0.1mol / L carbonate buffer containing 1% trichloroacetic acid (including

[0054] liquid), and added to the enzyme label assay wells that had been irradiated with a germicidal ultraviolet lamp for 30 minutes in advance;

[0055] (b) Warm bath at 37°C for 1 hour;

[0056] (c) Place at 4°C for 12 hours;

[0057] (d) Dry the solution, add skimmed milk powder or albumin blocking solution, and seal in a warm bath at 37°C;

[0058] (e) Dry the solution, wash with the washing solution three times, and store at 4°C.

[0059] kit includes

[0060] (a) Bacterial LPS monoclonal antibody preserved in 0.1% sodium azide, 50% glycerol disodium hydrogen phosphate solution;

[0061](b) immunoglobulin of peroxidase-labeled anti-LPS monoclonal...

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Abstract

The encapsulation process of bacterial lipopolysaccharide (LPS) includes dissolving LPS in 0.1 mol/L carbonate buffering liquid of pH 9.6 and containing 0.02-2 % trichloroacetic acid and adding into enzyme labelling detecting hole irradiated with ultraviolet ray in advance, treating in 35-39 deg.c warming bath; setting at 4 deg.c, spin drying and confining in 35-39 deg.c warming bath by adding defatted milk powder or albumin as confining liquid; and spin drying, washing and maintaining at 4 deg.c. Furthermore, reagent kit for detecting specific bacterial LPS in the the said method and the fast detection method are also provided. The reagent kit includes: bacterial LPS monoclonal antibody; second enzyme labelling antibody; standard LPS; diluent liquid; and encapsulated enzyme labelling detecting strip. The present invention has the advantages of being simple and convenient.

Description

technical field [0001] The present invention relates to a coating method of bacterial lipopolysaccharide (lipopolysaccharide, LPS), and further relates to a kit for detecting bacterial specific lipopolysaccharide provided by the method and a method for rapidly detecting bacterial specific LPS using the kit. Background technique [0002] Lipopolysaccharide is the main component of the cell wall of Gram-negative bacteria. As an endotoxin, it is also one of the important pathogenic factors of pathogenic bacteria. LPS has both positive and negative effects on the animal body. On the one hand, LPS brings toxic effects such as pyrogen reaction to the host infected by the bacteria, and makes the pathogenic bacteria avoid the attack of the animal complement, so that the pathogenic bacteria can successfully infect the host. On the other hand, , LPS is also an immunostimulant for the animal body, which can enhance the body's immune resistance. Since the discovery of endotoxin in the ...

Claims

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Application Information

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IPC IPC(8): C12Q1/02G01N33/53G01N33/577
Inventor 陈晓燕胡超群任春华
Owner SOUTH CHINA SEA INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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