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

Enzyme linked immunosorbent assay kit for detecting coumarin and indandione rodenticide and preparation and application thereof

An enzyme-linked immunosorbent reagent, indandione technology, applied in biological testing, measuring devices, material testing products, etc., can solve the problems of specificity, poor sensitivity, complicated operation, expensive instruments, etc., and achieve strong specificity and sensitivity. High, easy-to-use effects

Active Publication Date: 2020-06-16
北京维德维康生物技术有限公司
View PDF5 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

With the large-scale application of such rodenticides, rat poisons will inevitably remain in food, affecting human health
In recent years, poisoning incidents caused by such rodenticides have occurred frequently, seriously endangering people's health and life safety
[0003] At present, the detection methods for the above-mentioned anticoagulant rodenticides mainly include high-performance liquid chromatography and liquid chromatography-mass spectrometry. These methods have strong specificity and high sensitivity, but the operation is cumbersome and the equipment is expensive, so they are not suitable for the screening of large-scale samples. detection
The rapid detection method is mainly chemical colorimetric detection method, which has poor specificity and sensitivity, is prone to misjudgment, and cannot meet the needs of on-site detection.

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
  • Enzyme linked immunosorbent assay kit for detecting coumarin and indandione rodenticide and preparation and application thereof
  • Enzyme linked immunosorbent assay kit for detecting coumarin and indandione rodenticide and preparation and application thereof
  • Enzyme linked immunosorbent assay kit for detecting coumarin and indandione rodenticide and preparation and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] Embodiment 1, prepare the enzyme-linked immunosorbent assay kit that detects mouse Deke

[0048] The kit is composed as follows: ELISA plate (coated with the conjugate of mouse deke drug and carrier protein), antibody working solution (containing mouse deke drug monoclonal antibody), enzyme marker working solution (containing horseradish peroxidized Antibodies labeled with anti-modecyl monoclonal antibodies), washing solution, sample diluent, sample extract, working solution containing different gradient concentrations of mudecamin standards, substrate chromogenic solution, and stop solution.

[0049] The preparation method is as follows:

[0050] 1. Preparation of ELISA plate

[0051] 1. Preparation of Shudeke coating agent

[0052] (1) Preparation of Rat Deke Hapten

[0053] Weigh 100 mg of the raw material of Shudeke, dissolve it in 10 ml of methanol, add 30 mg of sodium hydride, and 36 mg of tetrabromobutyric acid to react at room temperature, after TLC detects t...

Embodiment 2

[0096] The using method of embodiment 2, embodiment 1 kit

[0097] 1. Sample pretreatment

[0098] 1. Pretreatment of meat samples, dairy products and serum samples

[0099] Accurately weigh 1 ± 0.01 g (or mL) of sample and add 5 mL of sample extract, at room temperature (25 ± 2°C), vortex at high speed for 1 min, and centrifuge at 4000 g for 5 min to obtain a sample solution. Take 200 μL sample solution and add 200 μL sample diluent, vortex at high speed for 1 min, and take 50 μL supernatant for analysis.

[0100] Two, use embodiment 1 kit to detect

[0101] 1. Take the microplate and insert it into the microplate rack, and record the position of each standard and sample, make 3 parallels for each sample, seal the unused microplate strip with a ziplock bag, and store it in a 2- 8°C environment;

[0102] 2. Add 50 μL of each standard working solution or sample solution into the corresponding standard or sample well;

[0103] 3. Add 50 μL antibody working solution to each ...

Embodiment 3

[0125] Specificity, detection limit, accuracy, precision detection of embodiment 3, embodiment 1 kit

[0126] 1. Specificity test of the kit:

[0127] The specificity of the Mouse Deke ELISA Kit is determined by cross-reaction tests with corresponding substances.

[0128] Ratac and commonly used rat poisons (bromadiolone, Dalong, warfarin, chlorwarfarin, chlordiquat, chlordiquat, fludifarin, grammide, coumatetyl, and rodentone) were prepared separately. Serial dilutions were performed according to Example 2, and the "Mutac standard working solution" was replaced with the serial dilutions of Mudak and its analogues to make a standard curve and find out the respective 50% inhibition on the curve. Concentration (IC 50 ), the specific method is as follows: get the mouse deg concentration (μg / L) corresponding to the ordinate value equal to 50%, that is, IC 50 value. Use the following formula to calculate the cross-reactivity rate of the kit to Ratac and various commonly used ra...

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 discloses an enzyme linked immunosorbent assay kit for detecting coumarin and indandione rodenticide and preparation and application thereof. The kit provided by the invention is composed of an elisa plate, an antibody working solution, an enzyme marker working solution, a washing solution, a sample diluent, a sample extracting solution, a solution containing murich standard substances with different gradient concentrations, a substrate developing solution and a stop solution. The murine decapeptide detection kit provided by the invention can be used for detecting coumarin and indandione rodenticide in food and human serum; in pork, milk and serum, the detection limit of murine, clorfenapyr and diphacinone is 5.0 [mu]g / kg, the detection limit of clorfenapyr is 2.5 [mu]g / kg, the detection limit of bromadiolone, dazomet and warfarin is 10.0 [mu]g / kg, the detection limit of trifluralin is 15.0 [mu]g / kg, and the detection limit of chlorfenapyr is 5.0 [mu]g / kg; the inter-batchvariable coefficient is less than 10%, and the intra-batch variable coefficient is less than 15%. The kit has the advantages of being easy and rapid to operate, high in sensitivity, high in specificity and high in accuracy and has great value in rapid detection of food poisoning.

Description

technical field [0001] The invention belongs to the field of rapid detection of drug residues, and relates to a kit for detecting coumarin and indanedione rodenticides, a preparation method and application thereof. Background technique [0002] There are many kinds of rodenticides, and anticoagulant rodenticides are the most widely used type of rodenticides, including coumarins such as coumarin, chlorwarfarin, fludifafarin, coumatemide, etc., and indane Diketones, such as Dalong, Diratide, and Chlorratine, competitively inhibit the action of vitamin K, inhibit the production of prothrombin and coagulation factors in the liver, increase the permeability and fragility of capillaries, and cause the death of rats from internal hemorrhage. With the large-scale application of such rodenticides, rat poisons inevitably remain in food, affecting human health. In recent years, poisoning incidents caused by such rodenticides have occurred frequently, seriously endangering people's hea...

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
Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/58G01N33/577G01N33/543G01N33/53
CPCG01N33/581G01N33/577G01N33/54306G01N33/5308Y02A50/30
Inventor 刘河冰温凯马立才杨柳刘薇崔乃元聂靖东邢维维丁亚芳
Owner 北京维德维康生物技术有限公司
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

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com