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

ELISA kit for detecting estriol and its application

A technology of triol enzyme linkage and estriol, which is applied in biological testing, measuring devices, material inspection products, etc., can solve the problems of abuse of antibiotics, achieve high accuracy, low requirements, and simple sample pretreatment process

Active Publication Date: 2018-07-06
BEIJING KWINBON BIOTECH
View PDF4 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is a phenomenon of abuse of antibiotics and hormones in the process of raising animals, resulting in a certain amount of estriol hormones in animal-derived food and water quality

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
  • ELISA kit for detecting estriol and its application
  • ELISA kit for detecting estriol and its application
  • ELISA kit for detecting estriol and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] The preparation of embodiment 1 kit components

[0028] 1. Preparation of estriol hapten

[0029] Take 1 g of estriol, add 20 ml of trifluoroacetic acid to dissolve, add 1.46 g of urotropine, heat, reflux for 3 h, cool to room temperature, add 30 ml of 1 mol / L dilute hydrochloric acid, and stir for 2 h. Stop the reaction, add water, adjust the pH value to 7 with sodium hydroxide, add ethyl acetate, extract, wash with water, dry and evaporate to dryness with anhydrous sodium sulfate, put on a silica gel column, petroleum ether / ethyl acetate (3 / 1, v / v) , to obtain hapten product 0.84g, yield 75.57%.

[0030] 2. Antigen preparation

[0031] Immunogen preparation—Estriol hapten was coupled with bovine serum albumin (BSA) to obtain immunogen.

[0032] Take 12 mg of aldehyde estriol hapten and dissolve it in 0.3 mL of DMF to obtain liquid A. Weigh 50 mg of BSA, fully dissolve in 4 mL of CB (pH 9.5), slowly add the reaction solution A to the protein solution drop by drop, ...

Embodiment 2

[0043] Embodiment 2 detects the formation of the ELISA kit of estriol

[0044] Set up the ELISA kit for detecting estriol to include the following components:

[0045] (1) A microtiter plate coated with an estriol-coupled antigen;

[0046](2) 5 bottles of estriol standard solution, the concentrations are 0μg / L, 0.05μg / L, 0.15μg / L, 0.45μg / L, 1.35μg / L;

[0047] (3) estriol secondary antibody labeled with horseradish peroxidase;

[0048] (4) Substrate chromogenic solution is made up of A liquid and B liquid, and A liquid is carbamide peroxide, and B liquid is tetramethylbenzidine;

[0049] (5) The stop solution is 2mol / L sulfuric acid;

[0050] (6) The washing solution has a pH value of 7.4, contains 0.5% to 1.0% Tween-20, 0.01‰ to 0.03‰ sodium azide preservative, and 0.1 to 0.3mol / L phosphate buffer, and the percentage is weight volume percentage;

[0051] (7) The complex solution is a phosphate buffer solution with a pH value of 7.0 and 0.02 mol / L, and the percentages are ...

Embodiment 3

[0052] The detection of estriol in embodiment 3 animal origin food, water quality

[0053] 1. Sample pretreatment

[0054] Animal tissue (chicken, pork, fish, shrimp) sample pretreatment method: Homogenize the tissue sample with a homogenizer; weigh 1.0±0.05g of the homogenized sample into a 50ml polystyrene centrifuge tube, add 5ml of ethyl acetate Esters, shake with a shaker for 1min, centrifuge at 3000g room temperature (20-25℃ / 68-77℉) for 5min; pipette 1ml of the upper organic phase into a 10ml clean and dry glass tube, place in a 50-60℃ (122-140℉) water bath Blow dry under nitrogen flow, add 1ml of n-hexane, 1ml of reconstituted working solution, vortex for 1min, mix well, centrifuge at 3000g room temperature (20-25℃ / 68-77℉) for 3min; remove the upper organic phase and the lower layer of water Phase 50 μl was used for analysis.

[0055] Milk sample: pipette 1ml of milk sample into a 50ml polystyrene centrifuge tube, add 5ml of ethyl acetate, shake with a shaker for 1min...

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

PropertyMeasurementUnit
recovery rateaaaaaaaaaa
Login to View More

Abstract

The invention provides an ELISA kit for detecting E3. The ELISA kit for detecting E3 comprises an enzyme label plate coated with a coating antigen, an E3 standardized product solution, an enzyme labeled secondary antibody, a substrate chromogenic solution, a stopping solution, washing liquid and a compound solution, the coating antigen is an E3 conjugated antigen, and the enzyme labeled secondary antibody is an enzyme labeled E3 secondary antibody. The invention further discloses a method for detecting E3 by using the ELISA kit, comprising the following steps: firstly, carrying out pretreatment on a sample and then detecting by using the kit, finally, analyzing detection results. The ELISA kit provided by the invention can be used for detecting the content of E3 in animal derived food and water samples, and is simple and convenient to operate, low in cost, high in sensitivity and capable of carrying out on-site monitoring and suitable for screening a large number of the samples.

Description

technical field [0001] The invention relates to an enzyme-linked immunoassay detection technology, in particular to an enzyme-linked immunoassay kit for detecting estriol, which can qualitatively and quantitatively detect the residual amount of estriol in animal-derived food and water quality. Background technique [0002] Estriol (E3) is a metabolite of estradiol and estrone, its value is very low during non-pregnancy, and it is mainly synthesized by the liver; during pregnancy, it is mainly synthesized by the fetal placenta, and after synthesis, it passes through the maternal blood circulation and is metabolized in the liver, and Sulfuric acid or glucuronic acid is combined to form conjugated E3, which is excreted in urine. E3 plays an important role in the regulation of fetal intrauterine development and can affect the sensitivity of the pregnant uterus to oxytocin. [0003] Animal-derived food and water quality are one of the important foods for human beings. They are r...

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 Patents(China)
IPC IPC(8): G01N33/74
CPCG01N33/74G01N2333/575
Inventor 罗晓琴韩深何方洋贾芳芳崔海峰冯才伟张瑜王兆芹杨春艳
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

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