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Method for detecting colimycin and special ELISA kit thereof

An enzyme-linked immunosorbent reagent and a technology for colistin, which is applied in the field of detecting colistin, can solve the problems that the detection sensitivity and detection limit cannot be achieved by using the instrument method, the thermal stability is poor, and it is not suitable for use, so as to shorten the detection time and cost. Low cost and low pre-treatment requirements

Inactive Publication Date: 2009-02-04
BEIJING WANGER BIOTECH
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

[0004] At present, the analytical method used to detect colistin residues is mainly microbial assay, but colistin is a high molecular weight (compared to commonly used antibiotics), high polar substance, and its poor thermal stability makes it unsuitable to use GC method analysis, its poor selectivity prevents the use of instrumental methods from achieving the required detection sensitivity and detection limit

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  • Method for detecting colimycin and special ELISA kit thereof
  • Method for detecting colimycin and special ELISA kit thereof
  • Method for detecting colimycin and special ELISA kit thereof

Examples

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Embodiment 1

[0032] Example 1. Preparation and use of a kit using the conjugate of colistin hapten and carrier protein as the coating source and enzyme-labeled secondary antibody as the enzyme label

[0033] 1. The detection principle of the kit using the conjugate of colistin hapten and carrier protein as the coating source and enzyme-labeled secondary antibody as the enzyme label is as follows:

[0034] When the coating on the microwell strip of the microtiter plate was originally a conjugate of colistin hapten and carrier protein, add the standard solution or sample solution to the microwell of the microtiter plate, and then add the specific colistin Antibody, the residual colistin in the sample competes with the colistin coupling antigen on the enzyme-labeled plate for the colistin specific antibody, and then the enzyme-labeled anti-antibody is added, and the color is developed with the color developing solution. The content is negatively correlated, and the residual content of colistin in...

Embodiment 2

[0131] Example 2. The kits for detecting colistin can also be as follows:

[0132] 1. A kit in which the coating source is a specific antibody and the enzyme label is an enzyme-labeled colistin hapten

[0133] (1) The working principle of this kit is:

[0134] When pre-coating the colistin-specific antibody on the microwell strip, add the sample solution or standard solution, and then add the enzyme-labeled colistin hapten solution. The colistin or colistin standard in the sample competes with the enzyme-labeled antigen. The colistin-specific antibody coated on the ELISA plate is used to develop the color. The sample absorbance value and the colistin content in the sample It is negatively correlated, and the content of colistin in the sample can be obtained by comparing with the standard curve. At the same time, the concentration range of colistin in the sample can be roughly judged by comparing the color of the colistin standard solution with a series of concentrations according ...

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Abstract

The present invention discloses a method of detecting colistin, and a special enzyme linked immunosorbent kit thereof. The enzyme linked immunosorbent kit comprises the colistin and specific antibodies of the colistin; and the specific antibodies are polyclonal antibodies or monoclonal antibodies of the colistin. The kit adopts the high specificity monoclonal antibodies of the colistin, which guarantees the reliability of detection results, and the experimental results show that the kit has the characteristics of high specificity, high sensitivity, high precision, high accuracy and the like; and the main reagents of the kit all adopt the form of working liquid, the operation is convenient, and the cost is low. The operation of the method which adopts the kit to detect the colistin is simple, the detection time is shortened, the operation have low requirements for the pretreatment of the sample, and the method can be used for simultaneously and rapidly detecting the samples in bulk. Therefore, the method which adopts the enzyme linked immunosorbent kit for detection can play a significant role in the detection of the colistin.

Description

Technical field [0001] The invention relates to a method for detecting colistin and a special enzyme-linked immunoassay kit. Background technique [0002] Colistin (colistin, CLS), also known as polymyxin E (polymyxin E), is produced by multiple colistin (B.polymyxa). It has a strong inhibitory effect on gram-negative bacteria. It can treat infections caused by Shigella dysentery, Escherichia coli, Pseudomonas aeruginosa, Salmonella and Proteus vulgaris. It can also be used as a feed additive. Improve the feed conversion rate, promote the growth of livestock and poultry, and prevent the occurrence of livestock and poultry diseases in large-scale breeding. Because colistin sulfate has strong antibacterial effects, low residue levels, and resistance to resistance, many countries and regions such as China, the United States, the European Union, and Japan have approved it as a feed additive or veterinary drug. [0003] With the increasing application of colistin sulfate in the aquacu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/53G01N33/577G01N33/535C12N5/12
Inventor 沈建忠何方洋万宇平冯才伟汪善良刘福林赵正苗冯才茂余厚美
Owner BEIJING WANGER BIOTECH
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