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

Time-resolved fluorescent immunoassay test strip for quantitative determination of tetracycline and norfloxacin

A technology of time-resolved fluorescence and norfloxacin, applied in the field of immunological detection, can solve the problems of tetracycline residue, detection cost cannot be expanded, and high sensitivity

Active Publication Date: 2020-01-24
南京海关动植物与食品检测中心
View PDF7 Cites 3 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, tetracycline is easy to remain in livestock and poultry, and then endanger human health through the food chain
Tetracycline antibiotics will bind to calcium ions in bones, affecting the development of teeth and bones, and can also cause damage to the digestive tract, liver and kidneys. Humans often have great risks if taken in large amounts
The detection methods involved in GC method and HPLC method have high sensitivity but require relatively expensive instruments and detection costs cannot expand their application
ELISA method, GICA method and other methods are relatively simple and convenient to operate, but have poor stability and low repeatability

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
  • Time-resolved fluorescent immunoassay test strip for quantitative determination of tetracycline and norfloxacin
  • Time-resolved fluorescent immunoassay test strip for quantitative determination of tetracycline and norfloxacin
  • Time-resolved fluorescent immunoassay test strip for quantitative determination of tetracycline and norfloxacin

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0036] In the present invention, the preparation method of the time-resolved fluorescent immunoassay strip for quantitative determination of tetracycline and norfloxacin preferably comprises:

[0037] Soak the glass cellulose membrane in the conjugation pad treatment solution, take it out and dry it to obtain a dry membrane, and spray the Eu-tetracycline monoclonal antibody and Eu-norfloxacin monoclonal antibody labeled with the Eu-time-resolved fluorescent nanospheres On the dry film, obtain the bonding pad;

[0038] Soak the glass cellulose membrane in the sample pad treatment solution, take it out and dry it to obtain the sample pad;

[0039] Draw the first test line, the second test line and one quality control line on the nitrocellulose membrane, the first test line is coated with tetracycline artificial antigen or norfloxacin artificial antigen, and the second test line is coated with norfloxacin Sandacin artificial antigen or tetracycline artificial antigen is coated w...

Embodiment 1

[0046] Artificial antigen preparation:

[0047] Tetracycline (TC) and norfloxacin (NFLX) drugs were coupled to carrier protein by diazo method. The artificial antigen preparation is shown in Table 1 below:

[0048] Table 1 Artificial antigens

[0049]

[0050] 1. Solution preparation

[0051] (a) NaOH solution: the concentration is 0.2mol / L;

[0052] (b)NaNO 2 Solution: mass fraction is 1%;

[0053] (c) HCl solution: the concentration is 0.2mol / L;

[0054] (d) BSA solution: 100mg BSA dissolved in 10mL 0.01mol / L PBS (pH 7.2-7.6);

[0055] (e) HSA solution: 50 mg of HSA was dissolved in 10 mL of 0.01 mol / L PBS (pH 7.2-7.6).

[0056] 2. Small molecule coupling carrier protein

[0057] (1) Weigh an appropriate amount of tetracycline (TC) and norfloxacin (NFLX) and dissolve them in 1 mL (a) (equimolar amounts);

[0058] (2) Add 500 μL of (b) to the tetracycline (TC) and norfloxacin (NFLX) drug solution in (1), and pre-cool in a water bath at 4°C;

[0059] (3) Add the s...

Embodiment 2

[0065] Antibody preparation

[0066] 1. Animal immunity:

[0067] Female Balb / c mice aged 6-8 weeks were selected as immunization objects. Before the initial immunization, blood was collected from the tail vein of each mouse, and 200 μL of the emulsified immunogen (artificial antigen A plus complete adjuvant) was injected intraperitoneally during the initial immunization; On the 14th day and 28th day after immunization, the second and third immunizations (artificial antigen A plus incomplete adjuvant) were carried out in the same way, and the orbital blood was collected from each mouse on the 5th day after the third immunization. Serum was used to evaluate the antibody titer. Booster immunization was carried out on the seventh day after the third immunization, and 100 μL of immunogen was injected into the tail vein. On the third day after the booster immunization, splenic lymphocytes and Sp2 / 0 (mixing ratio of 5:1) were collected for cell Fusion, fusion using the PEG method. ...

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
particle diameteraaaaaaaaaa
Login to View More

Abstract

The invention provides a time-resolved fluorescent immunoassay test strip for quantitative determination of tetracycline and norfloxacin, and belongs to the technical field of immunological detection.The test strip sequentially comprises a sample adding area, a binding area and a detection area, wherein the detection area comprises a first detection line, a second detection line and a quality control line in sequence; the first detection line is coated with a tetracycline artificial antigen or norfloxacin artificial antigen, the second detection line is coated with a norfloxacin artificial antigen or tetracycline artificial antigen, and the binding region is tiled with Eu-time-resolved fluorescent nanosphere-labeled antibody Eu-tetracycline monoclonal antibody and Eu-norfloxacin monoclonal antibody. The time-resolved fluorescent immunoassay test strip for the quantitative determination of tetracycline and norfloxacin can quickly quantify tetracycline and norfloxacin in samples with low detection limits, high sensitivity and good repeatability.

Description

technical field [0001] The invention belongs to the technical field of immunological detection, and in particular relates to a time-resolved fluorescent immunological test strip for quantitative determination of tetracycline and norfloxacin. Background technique [0002] Tetracycline (TC) is a class of broad-spectrum antibiotics, which mainly achieve antibacterial effect by inhibiting bacterial protein synthesis, and are widely used in livestock and poultry production. However, tetracycline is easy to remain in livestock and poultry, and then endanger human health through the food chain. Tetracycline antibiotics will bind to calcium ions in bones, affecting the development of teeth and bones, and can also cause damage to the digestive tract, liver and kidneys. Humans often have great risks if taken in large amounts. Tetracycline (tetracycline, TC), chlortetracycline (chlortetracycline, CTC), oxytetracycline (oxytetracycline, OTC), doxycycline (doxycycline, DC), etc. Ring b...

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): G01N33/543G01N33/58G01N33/558G01N33/577
CPCG01N33/54326G01N33/582G01N33/587G01N33/558G01N33/577Y02A50/30
Inventor 王毅谦龙云凤高玲陈雷张晓燕柳菡李静静沈伟健
Owner 南京海关动植物与食品检测中心
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