Method and test strip for detecting zilpaterol, and application of test strip

A detection line and reaction technology, which is applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of lagging and hysteresis of detection methods, and achieve the effects of short detection time, strong specificity and high sensitivity

Inactive Publication Date: 2016-02-17
JINAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the government explicitly prohibits the use of clenbuterol, poisoning incidents still occur from time to time. One of the important re

Method used

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  • Method and test strip for detecting zilpaterol, and application of test strip
  • Method and test strip for detecting zilpaterol, and application of test strip
  • Method and test strip for detecting zilpaterol, and application of test strip

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0067] The preparation of embodiment 1 zilpaterol monoclonal antibody

[0068] (1) Basic test steps for zilpaterol antigen synthesis

[0069] ① Accurately weigh 22 mg of zilpaterol and dissolve it in 2 ml of ice-cold hydrochloric acid solution of pH 3.0 and 0.1 mol / L. Slowly add 2 mL of ice-cold sodium nitrite solution with a pH of 2.0 and 30 mg / mL, and carry out diazotization reaction at 0-5°C for 20 min to obtain zilpaterol diazonium salt. At the same time, 100mg of bovine serum albumin was accurately weighed and dissolved in 5ml of pH8.6, 0.1mol / L phosphate buffer solution, and the reacted zilpaterol diazonium salt was added dropwise, adjusted with 1mol / L sodium hydroxide solution. The pH value was adjusted to 7.5 and reacted overnight at 4°C. The reaction solution was dialyzed with pH 7.4, 0.01 mol / L PBS solution (containing 0.15 mol / L NaCl) for 3 days, the water was changed 4 times a day, and the dialyzed reaction solution was collected to obtain zilpaterol antigen. Th...

Embodiment 2

[0082] The preparation of embodiment 2 binding pad

[0083] 1. Preparation of colloidal gold

[0084](1) Colloidal gold with an average particle size of about 20, 30, and 40 nm was respectively prepared by the trisodium citrate method. The preparation method of 20nm colloidal gold is as follows: Utilize the condensing reflux device to heat 50mL of chloroauric acid solution with a mass fraction of 0.01% to boiling, then quickly add 1.2ml of trisodium citrate solution with a mass fraction of 1%, and continue to boil after the color is stable 10min, cooled, and stored at 4°C for later use. Colloidal gold of 30 and 40 nm was prepared in the same way, except that 0.75 and 0.5 ml of 1% trisodium citrate solution were added respectively. The distribution and morphology of the three kinds of colloidal gold were observed by spectral scanning, nano-force laser and transmission electron microscope. After testing, colloidal gold with average particle diameters of 20, 29, and 40 nm were...

Embodiment 3

[0092] Embodiment 3 detects the preparation of test paper

[0093] 1. Materials required for the production of test strips

[0094] ①Nitrocellulose membrane (NC membrane): imported from Millipore / NC of the United States, the specification of nitrocellulose membrane is 30cm×3m / HAHY00010:

[0095] The area of ​​nitrocellulose membrane required for one immunofluorescence test strip is: 2.5cm×0.6cm.

[0096] The total area of ​​nitrocellulose membrane required to produce 100,000 test strips is:

[0097] 2.5cm×0.6cm×100,000=1.56×10 5 cm 2 ;

[0098] The total number of rolls of nitrocellulose membrane required to produce 100,000 test strips:

[0099] 2.25×10 5 cm 2 / 30×300cm 2 = 20 volumes.

[0100] ②The total amount of antigen required: 4mg / mL×2.0μl / cm×0.25cm×100,000=20mg

[0101] ③The total amount of goat anti-rabbit IgG required: 1mg / ml×2.0μl / cm×0.25cm×100,000=50mg

[0102] 2. Spraying of C (quality control line) and T (test line) on the fiber membrane

[0103] The s...

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Abstract

The invention discloses a method and test strip for detecting zilpaterol, and application of the test strip. The method is a combination of colloidal-gold immunochromatography assay and competitive immunochromatography assay; an antibody used in the method is a colloidal-gold-labeled monoclonal antibody against zilpaterol, and competitive antigen zilpaterol is immobilized on a detection line of the test strip; a to-be-tested sample reacts with the colloidal-gold-labeled monoclonal antibody against zilpaterol at first and then with the competitive antigen; when a red strip does not appear on the detection line, it is determined that the to-be-tested sample contains zilpaterol; and when a red strip appears on the detection line, it is determined that the to-be-tested sample does not contain zilpaterol or has low zilpaterol content which does not exceed detection limit. The test strip comprises a base plate, and a sample absorption pad, a bonding pad, a chromatographic membrane and a water absorbing pad which are all adhered on the base plate and closely connected in sequence, wherein the bonding pad is provided with the colloidal-gold antibody, and the chromatographic membrane is provided with a detection line, a quality control line and the detection line. When the test strip is used to detect a positive sample, no color is displayed on the detection line.

Description

technical field [0001] The invention belongs to the field of food safety detection and the technical field of immune detection, in particular to a method for detecting zilpaterol, a test strip and an application thereof. Background technique [0002] Food safety is a major livelihood issue, which is related to the health and life safety of the general public, and is also an important foundation for national economic development and social stability. In recent years, various problems of low food safety in my country have had a negative impact on the public, society, economy, local government image, and my country's international reputation. The monitoring of food safety and the development of detection technology need to be paid special attention to. [0003] Zipaterol is a β2 adrenergic receptor agonist, which can selectively stimulate the β2 receptors of smooth muscle, so that the nutrients in various animals such as cattle, pigs, sheep and poultry are transferred from fat ...

Claims

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Application Information

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IPC IPC(8): G01N33/577
Inventor 朱伟杰刘红
Owner JINAN UNIVERSITY
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