Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Magnetic immunochemiluminescence detection kit for apramycin, and applications thereof

A chemiluminescent detection and apramycin technology, which is applied in the field of immunological detection, can solve the problems of low sensitivity, high false negative rate and false positive rate, and achieve high sensitivity and specificity, short detection time, and fully automated Effect

Active Publication Date: 2018-08-03
BEIJING KWINBON BIOTECH
View PDF4 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the most commonly used immunoassay methods include enzyme-linked immunoassay and colloidal gold immunochromatographic assay, but due to their low sensitivity, high false negative rate and false positive rate, they are gradually replaced by high sensitivity, high accuracy and short detection time. short chemiluminescent immunoassay

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Magnetic immunochemiluminescence detection kit for apramycin, and applications thereof
  • Magnetic immunochemiluminescence detection kit for apramycin, and applications thereof
  • Magnetic immunochemiluminescence detection kit for apramycin, and applications thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Example 1: Preparation of Specific Components of Apramycin Magnetic Immunochemiluminescent Detection Kit

[0031] 1. Synthesis of Apramycin hapten (see attached for synthetic route figure 1 )

[0032] A. Take 1.0g of apramycin, add 50mL of pyridine to dissolve, add 0.83g of pyridinium dichromate (PDC), stir at 80°C for 3h, stop the reaction, evaporate to dryness, add toluene, evaporate to dryness, and get black The oil was put on a silica gel column and eluted with dichloromethane / methanol (V / V, 5 / 1) to obtain 0.89 g of aldehyde apramycin, with a yield of 89.35%;

[0033] B. Take 0.89g of alpramycin, add 80mL of methanol to dissolve, add 0.32g of adipic acid 1-hydrazide, add 0.3g of sodium carbonate, stir at room temperature for 2h, stop the reaction, rotary steam, remove methanol, add water, ethyl acetate The ester was extracted and shaken, evaporated to dryness by rotary evaporation, and recrystallized from dichloromethane / methanol (V / V, 1:1) to obtain 0.71 g of apr...

Embodiment 2

[0054] Example 2: Construction of Apramycin Magnetic Immunochemiluminescence Detection Kit

[0055] The Apramycin Magnetic Immunochemiluminescence Detection Kit was assembled to include the following components:

[0056] (1) horseradish peroxidase-labeled apramycin hapten;

[0057] (2) The enzyme-labeled antigen diluent is pH 7.2-7.6, Na 3 PO 4 A buffer solution with a concentration of 0.01mol / L and a NaCl concentration of 0.25mol / L;

[0058] (3) Apramycin monoclonal antibody labeled with immunomagnetic beads;

[0059] (4) Magnetic label antibody diluent is pH 7.2-7.6, Na 3 PO 4 A buffer solution with a concentration of 0.01mol / L and a NaCl concentration of 0.25mol / L;

[0060] (5) Apramycin series standard solution concentrations are 0μg / L, 0.3μg / L, 0.9μg / L, 2.7μg / L, 5.4μg / L, 10.8μg / L;

[0061] (6) The chemiluminescence substrate A solution is trishydroxymethylaminomethane solution containing luminol and p-cresol, and the chemiluminescence substrate B solution contains ...

Embodiment 3

[0064] Embodiment 3: the detection of apramycin residual amount in the sample

[0065] 1. Sample pretreatment

[0066] Add 30 μL milk sample to 870 μL complex solution and vortex for 1 min for detection.

[0067] 2. Detection with kit

[0068] 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 magnetic label antibody working solution container of the chemiluminescence detector; set the position on the sample rack for each sample / standard product, enter the sample information and the name of the test item to be tested ;Put the sample tube / standard tube into the set sample rack, and the chemiluminescence detector draws 50 μL enzyme-labeled antigen working solution, 50 μL sa...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention provides a magnetic immunochemiluminescence detection kit for apramycin. The magnetic immunochemiluminescence detection kit comprises an enzyme labeled antigen, an enzyme labeled antigendilution solution, a magnetically labeled antibody, a magnetically labeled antibody dilution solution, apramycin series standard solutions, a chemiluminescent substrate solution A, a chemiluminescentsubstrate solution B, a redissolution solution and a washing solution, wherein the enzyme labeled antigen is horseradish peroxidase-labeled apramycin hapten, and the magnetically labeled antibody isimmunomagnetic bead labeled apramycin monoclonal antibody. The invention further discloses a method for detecting the residual amount of apramycin in milk by using a chemiluminescence detector supporting the kit. According to the present invention, the method has advantages of high sensitivity, high specificity and short detection time in the detection of apramycin.

Description

technical field [0001] The invention relates to a chemiluminescence detection kit and a detection method thereof, in particular to a magnetic immunochemiluminescence detection kit for detecting residual apramycin in milk, belonging to the field of immunological detection. Background technique [0002] Apramycin (Apramycin, Apra), also known as dark mycin, apramycin, abramycin, is a typical representative drug of aminoglycoside antibiotics. Apramycin is a new product developed by Eli Lilly and Company in 1985. It can also be produced in my country at present. It is produced by fermentation of a mutant strain of Streptomyces tenebrarius. Apramycin sulfate is mostly used in feed, which is brown crystalline powder or dull yellow powder with strong polarity, easily soluble in water and slightly soluble in ethanol. [0003] Improper use of aminoglycoside antibiotics can cause toxic side effects on the kidneys and hearing of animals. Therefore, the maximum residue limits of amino...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): G01N33/94
CPCG01N33/9446G01N2430/00
Inventor 万宇平何方洋刘清珺曹东山张瑜冯月君崔海峰宋灏
Owner BEIJING KWINBON BIOTECH
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products