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Lincomycin magnetic immunochemiluminescence detection kit and application thereof

A chemiluminescent detection and lincomycin technology, applied in the field of immunological detection, can solve the problems of unsuitable screening of a large number of samples, expensive, complicated and cumbersome operation processes, etc., to reduce human operation errors, short detection time, and realize fully automated effect

Active Publication Date: 2015-09-09
BEIJING KWINBON BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to expensive instruments and equipment, complex and cumbersome operating procedures, and high skill requirements for inspectors, the above-mentioned instrumental detection methods are not suitable for the screening of a large number of samples

Method used

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  • Lincomycin magnetic immunochemiluminescence detection kit and application thereof
  • Lincomycin magnetic immunochemiluminescence detection kit and application thereof
  • Lincomycin magnetic immunochemiluminescence detection kit and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Embodiment 1: Preparation of specific components of the kit

[0036] 1. Lincomycin Hapten Synthesis

[0037]Lincomycin hapten is ketolincomycin obtained by oxidizing the alcoholic hydroxyl group in lincomycin with pyridinium dichromate, and then condensing ketolincomycin with aminobutyric acid to obtain a four-carbon chain carboxyl hapten. Add 0.8g~1.2g lincomycin to acetonitrile to dissolve, then add 0.80g~0.85g pyridinium dichromate, stir, add 0.5mL~1.1mL acetic anhydride and 0.3mL~0.7mL acetic acid, stir at room temperature for 6h, After the reaction was complete, the reaction was stopped and evaporated to dryness, purified on a silica gel column, eluted and separated with ethyl acetate:petroleum ether=1:1, and the product 1 was obtained. Take the product 1 and dissolve it in methanol, then add 2mL-4mL of an aqueous solution containing 1.0g-1.4g of aminobutyric acid and 2mL of an aqueous solution of 0.5g-0.8g potassium hydroxide, and heat the reaction at 60°C for 7...

Embodiment 2

[0053] Embodiment 2: the formation of kit

[0054] A magnetic immunochemiluminescence detection kit for detecting lincomycin drugs was constructed so that it contained the following components:

[0055] horseradish peroxidase-labeled lincomycin hapten

[0056] Enzyme-labeled Antigen Diluent

[0057] Conjugates of lincomycin monoclonal antibody and magnetic beads

[0058] Magnetic Label Antibody Diluent

[0059] Lincomycin standard solution, the concentrations are: 0μg / L, 0.3μg / L, 0.9μg / L, 1.8μg / L, 5.4μg / L, 10.8μg / L, the standard diluent is pH7.4, Containing 0.05% Tween-20, 0.05mol / L phosphate buffer. The percentage content is a mass percentage content.

[0060] Concentrated complex solution is concentrated phosphate buffer solution, which contains 5.0g~8.0g NaH per liter 2 PO 4 2H 2 O, 30.0g ~ 35.0g Na 2 HPO 4 12H 2 Aqueous solution of O.

[0061] The concentrated washing solution is a phosphate buffer solution with a volume fraction of 0.03%-0.08% Tween-20, pH=7.4...

Embodiment 3

[0062] Embodiment 3: the detection of lincomycin residual amount in the sample

[0063] 1. Sample pretreatment method

[0064] (1) milk

[0065] Take 50 μL fresh milk sample and add 950 μL sample diluent (dilute the concentrated complex solution with deionized water at a volume ratio of 1:1), vortex and mix well, and take the solution for sample analysis.

[0066] (2) milk powder

[0067] Weigh 0.5g±0.05g milk powder sample, add 5mL sample diluent, vortex mix, take 200μL from it and add to 600μL sample diluent, vortex mix, take the solution for sample analysis.

[0068] 2. Detection with kit and result analysis

[0069] Dilute the enzyme-labeled antigen and the enzyme-labeled antigen diluent at a volume ratio of 1:10 to 1:20, and put it into the enzyme-labeled antigen working solution container of the chemiluminescent detector; dilute the magnetic-labeled antibody and the magnetic-labeled antibody diluent at 1:1 : Dilute at a volume ratio of 10 to 1:20, and put it into the...

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Abstract

The invention relates to a magnetic immunochemiluminescence kit for detecting lincomycin, which comprises the following reagents, enzyme-labelled antigen, an enzyme-labelled antigen weak solution, a magnetic antibody, a magnetic antibody weak solution, a lincomycin series standard substance solution, a chemiluminescence substrate solution A, a chemiluminescence substrate solution B, a concentrated compound solution, and a concentrated washing liquid. enzyme-labelled antigen is a horseradish peroxidase-labelled lincomycin hapten marker, lincomycin hapten is keto lincomycin obtained by performing pyridinium dichromate cas oxidation on alcoholic hydroxyl group from lincomycin, keto lincomycin and aminobutyric acid are subjected to a condensation reaction to obtain four-carbon chain carboxyl hapten, the magnetic antibody is obtained by coupling the lincomycin monoclonal antibody and magnetic bead. The invention also relates to a method for detecting lincomycin in milk and milk powder by employing the kit and a matched chemiluminescence detector, and the method has advantages of high sensitivity, specificity and short detection time for detecting lincomycin.

Description

technical field [0001] The invention relates to a chemiluminescence detection kit and a detection method thereof. Especially the magnetic immunochemiluminescence detection kit for detection of lincomycin residues in milk, milk powder and other samples. It belongs to the field of immunological detection. Background technique [0002] Lincomycin (Lincomycin, LIN), also known as lincomycin, is a lincosamide antibiotic produced by the fermentation of Streptomyces lincolinus. Lincomycin has strong antibacterial activity, mainly acts on Gram-positive bacteria, and is an RNA-dependent acetylcholinesterase inhibitor. As a feed additive, lincomycin has the effect of increasing the rate of weight gain and improving feed remuneration. The method of use is mainly to add lincomycin products and mix them in the feed, or directly inject into the udder of sick cows to treat cow mastitis. Both methods of administration will cause the drug to remain in the milk. According to literature re...

Claims

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Application Information

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IPC IPC(8): G01N33/577
CPCG01N33/54326
Inventor 何方洋冯才伟杜美红崔海峰聂雯莹徐念琴马腊腊何勇
Owner BEIJING KWINBON BIOTECH
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