Unlock instant, AI-driven research and patent intelligence for your innovation.

Magnetic particle chemiluminescence kit for detecting diazepam, and applications thereof

A technology of chemiluminescent reagents and kits, which is applied in the fields of chemiluminescence/bioluminescence, analysis through chemical reactions of materials, and measurement devices, which can solve the problems of cumbersome operation, high detection cost, and poor stability of kits, and achieve The effect of low detection time, fast detection and high sensitivity

Inactive Publication Date: 2013-02-13
BEIJING KWINBON BIOTECH
View PDF0 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

High-performance liquid chromatography is cumbersome to operate, expensive to detect, requires high testing personnel, and takes a long time, which is not conducive to popularization and application
Enzyme-linked immunosorbent assay detection requires multiple steps of plate washing, which is relatively time-consuming and difficult to improve the degree of automation of the operation. Enzyme-catalyzed reactions require high temperature and poor stability of the kit.

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 particle chemiluminescence kit for detecting diazepam, and applications thereof
  • Magnetic particle chemiluminescence kit for detecting diazepam, and applications thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] The preparation of embodiment 1 kit specific components

[0031] 1) Synthesis of stable hapten

[0032] The diazepam hapten is prepared by condensation of diazepam and carboxymethyl hydroxylamine.

[0033] 2) Preparation of stabilized artificial antigen

[0034] The immunogen is obtained by coupling the stable hapten with bovine serum albumin by a water-soluble carbodiimide method.

[0035] 3) Preparation of monoclonal antibodies

[0036] Animal immunization: Immunize Balb / c mice with the immunogen at a dose of 100 μg / mouse to produce antiserum.

[0037] Cell fusion and cloning: Splenocytes from immunized Balb / c mice were fused with SP2 / 0 myeloma cells at a ratio of 9:1 to obtain hybridoma cell lines of monoclonal antibodies.

[0038] Cell cryopreservation and recovery: the hybridoma cells were made into 1×10 cryopreservation medium 9 cells / ml for long-term storage in liquid nitrogen. When recovering, take out the cryopreservation tube, put it into a 37°C water ba...

Embodiment 2

[0049] The formation of embodiment two kits

[0050] Construct the direct competition chemiluminescent detection kit for detecting the residual magnetic particles of diazepam so that it contains the following components:

[0051] Fluorescent marker for FITC-labeled diazepam monoclonal antibody

[0052] Luminescent marker of ABEI-labeled stable hapten

[0053] Separation reagent of paramagnetic nanobeads coated with goat anti-FITC monoclonal antibody

[0054] Stability standard solution (0ng / ml, 0.01ng / ml, 0.03ng / ml, 0.1ng / ml, 0.3ng / ml, 1.0ng / ml), the standard diluent is pH7.2, 0.3%NaN 3 , 0.05mol / L TRIS buffer. The percentage content is a mass percentage content.

[0055] The concentrations of stable quality control solution are 0.02ng / ml and 0.8ng / ml respectively, and the dilution solution of quality control is pH7.4, 0.03% NaN3, 0.05mol / L TRIS buffer. The percentage content is a mass percentage content.

[0056] Concentrated lotion pH 7.2, 0.4% Tween-20, 0.3% NaN 3 , ...

Embodiment 3

[0057] The detection of stability in the actual sample of embodiment three

[0058] 1. Sample pretreatment

[0059] (1) Animal tissues (muscle, liver, etc.),

[0060] Take the tissue sample and homogenize it with a homogenizer above 4000rpm for 1min; weigh 2.0±0.05g of the homogenate into a 50ml centrifuge tube, add 8ml of 0.1M sodium hydroxide solution, and shake for 5min. Above 3000g, centrifuge at 15°C for 10min; take 1ml of the supernatant, add 10ml of n-hexane, shake for 5min, and above 3000g, centrifuge at 15°C for 5min; take 5ml of the upper n-hexane phase and dry it in a water bath at 50-60°C under nitrogen flow; add 1ml of complex Dissolve the working solution to dissolve the residue; take 50 μl for analysis, sample dilution factor: 10.

[0061] (2) Urine

[0062] Take 1ml of clear urine sample, add 4ml of 0.1M sodium hydroxide solution to a 50ml centrifuge tube, shake for 2-5min; take 1ml of the mixed solution into another centrifuge tube, add 10ml of n-hexane, sh...

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
Sensitivityaaaaaaaaaa
Login to View More

Abstract

The present invention relates to a magnetic particle chemiluminescence kit for detecting diazepam. The kit comprises a luminous marker, a fluorescein marker, a standard substance, a quality control substance and a separation reagent, wherein the luminous marker is isoluminol luminous marker labeled diazepam hapten, the fluorescein marker is fluorescein labeled diazepam monoclonal antibody or fluorescein derivative labeled diazepam monoclonal antibody, and the separation reagent is sheep anti-FITC monoclonal antibody coated paramagnetic nanometer microbeads. The present invention further relates to a method for detecting diazepam in foods of animal origin by using the kit, wherein the method provides characteristics of high sensitivity, high specificity and rapid detection for diazepam detection.

Description

technical field [0001] The invention relates to a chemiluminescent detection kit and a testing method thereof. Especially the magnetic particle chemiluminescence detection kit for detecting stability in food of animal origin. Background technique [0002] Diazepam, as a sedative, is widely used in livestock and poultry, which can reduce the amount of exercise of livestock and poultry animals, increase the weight of animals and ensure that livestock and poultry will not produce stress reactions during long-distance transportation. However, a large amount of diazepam taken by animals will lead to drug residues in food and be absorbed by the human body. Large doses can cause central nervous system depression, respiratory depression and other toxic manifestations. Long-term use of this type of drug can lead to dependence or addiction, and sudden withdrawal of the drug can cause withdrawal reactions, manifested as depression, agitation, and insomnia. [0003] At present, the me...

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): G01N21/76G01N33/577
Inventor 万宇平陈兰珍冯静杨秀贤聂雯莹陈炜玲
Owner BEIJING KWINBON BIOTECH