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Method for detecting colimycin and special ELISA kit thereof

An enzyme-linked immunosorbent reagent and colistin technology, which is applied in the field of colistin detection, can solve the problems that the detection sensitivity and detection limit cannot be achieved by using instrumental methods, poor thermal stability, and should not be used, so as to shorten the detection time and cost Low cost and low pre-treatment requirements

Inactive Publication Date: 2012-11-28
BEIJING WANGER BIOTECH
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  • Abstract
  • Description
  • Claims
  • Application Information

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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

Experimental program
Comparison scheme
Effect test

Embodiment 1

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

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

[0034] When the coating on the microwell strip of the microtiter plate is originally a conjugate of colistin hapten and carrier protein, add standard solution or sample solution to the microwell of the microtiter plate, and then add colistin-specific Antibody, the residual colistin in the sample competes with the colistin-coupled antigen on the ELISA plate for the colistin-specific antibody, then the enzyme-labeled anti-antibody is added, and the color is developed with a chromogenic solution. The absorbance value of the sample is consistent with that of the colistin The content is negative...

Embodiment 2

[0131] Embodiment 2, the kit for detecting colistin can also have the following several kinds:

[0132] 1. The original coating is a specific antibody, and the enzyme marker is an enzyme-labeled colistin hapten kit

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

[0134] When the colistin-specific antibody is pre-coated on the microwell strip, add the enzyme-labeled colistin hapten solution after adding the sample solution or standard solution. The colistin or colistin standard in the sample competes with the enzyme-labeled antigen for the colistin-specific antibody coated on the microtiter plate, and the color is developed with a chromogenic solution, and the absorbance value of the sample is related to the colistin content in the sample A negative correlation is formed, 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 ...

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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 ELISA kit. Background technique [0002] 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 and can treat infections caused by Shigella dysenteriae, Escherichia coli, Pseudomonas aeruginosa, Salmonella and Proteus vulgaris, and 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 farming. Because colistin sulfate has the characteristics of strong antibacterial effect, low residue, and not easy to produce drug 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...

Claims

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Application Information

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Patent Type & Authority Patents(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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