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Fluorescent microsphere immunochromatography test paper strip for detecting clenbuterol hydrochloride residue, preparation method and applications thereof

A technology of clenbuterol hydrochloride and immunochromatographic test paper, which is applied in biological testing, measuring devices, analytical materials, etc., can solve the problems that are not applicable to large-throughput rapid screening and testing of grassroots enterprises and institutions, and the professional requirements of operators are relatively high. High, matrix effect background interference and other problems, to achieve the effect of increasing stability and fluorescence lifetime, increasing reaction sensitivity, and high sensitivity

Active Publication Date: 2015-08-19
BEIJING KWINBON BIOTECH
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, since the above methods require advanced detection equipment, expensive detection costs, cumbersome steps, time-consuming, and high professional requirements for operators, they are not suitable for large-throughput rapid screening detection in grassroots enterprises and institutions.
Colloidal gold immunochromatography has the advantages of fast detection speed, low price, and simple operation. It is currently the main monitoring method for detecting clenbuterol hydrochloride at home and abroad, but there are still some defects, such as poor stability, low sensitivity, Quantitative detection cannot be realized, the matrix effect is obvious, the background interference is large, and the color is single, it is difficult to realize multiple detection and joint detection

Method used

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  • Fluorescent microsphere immunochromatography test paper strip for detecting clenbuterol hydrochloride residue, preparation method and applications thereof
  • Fluorescent microsphere immunochromatography test paper strip for detecting clenbuterol hydrochloride residue, preparation method and applications thereof
  • Fluorescent microsphere immunochromatography test paper strip for detecting clenbuterol hydrochloride residue, preparation method and applications thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Example 1 Composition of Fluorescent Microsphere Immunochromatographic Test Strips for Detecting Clenbuterol Hydrochloride Residues

[0037] 1. Test strips ( figure 1 , figure 2 )

[0038] The test strip is composed of a bottom plate, a sample binding pad, a nitrocellulose membrane and an absorbent pad;

[0039] The sample binding pad 1, the nitrocellulose membrane 2 and the water-absorbing pad 3 are overlapped and pasted on the base plate 6 in sequence, the end of the sample-binding pad is connected to the beginning of the nitrocellulose membrane, and the end of the nitrocellulose membrane is connected to the water-absorbing pad. The start of the sample binding pad is aligned with the start of the bottom plate, and the end of the absorbent pad is aligned with the end of the bottom plate;

[0040] The nitrocellulose membrane is fixed with detection area 4 and quality control area 5, and the detection area is sprayed with clenbuterol hydrochloride hapten-carrier prot...

Embodiment 2

[0042] Example 2 Preparation of Fluorescent Microsphere Immunochromatographic Test Strips for Detecting Clenbuterol Hydrochloride Residues

[0043] The preparation method of the fluorescent microsphere immunochromatographic test strip for detecting residual clenbuterol mainly comprises the following steps:

[0044] 1) Preparation of the sample binding pad: label the anti-clenbuterol monoclonal antibody with commercially available fluorescent microspheres, and dilute it with a specific buffer system, soak the sample binding pad in the dilution buffer, and vacuum freeze prepared after drying;

[0045] 2) Preparation of nitrocellulose membrane: Spray the clenbuterol hydrochloride hapten-carrier protein conjugate on the detection area of ​​the nitrocellulose membrane to make the detection area; spray goat anti-mouse anti-antibody onto the nitrocellulose The scope of the quality control area on the plain film is made into a quality control area;

[0046] 3) Assembly and cutting: th...

Embodiment 3

[0089] Example 3 Application of Fluorescent Microsphere Immunochromatographic Test Strips for Detecting Clenbuterol Hydrochloride Residues

[0090] 1. Sample pretreatment

[0091] (1) Urine sample

[0092] Take a clear urine sample for direct measurement. The sample must be collected in a clean and dry plastic urine cup or glass container without any preservatives; Freeze at -20°C, avoid repeated freezing and thawing.

[0093] (2) Animal tissue samples

[0094] Take the fat-free animal tissue sample and homogenize it in a homogenizer at 10000r / min for 1min; weigh 2.0g±0.05g of the homogenized tissue sample into a 4mL polystyrene centrifuge tube, add 500μL sample extract (0.35mol / L of PB buffer), vortexed for 1min; placed in an 80°C water bath for 10min, centrifuged at 4000r / min for 5min, cooled to room temperature, and ready to be tested.

[0095] 2. Test with test strips

[0096] Draw 100 μL of the sample solution to be tested and drop it vertically into the sample hole...

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Abstract

The present invention discloses a fluorescent microsphere immunochromatography test paper strip for detecting clenbuterol hydrochloride residue, a preparation method and applications thereof. The test paper strip comprises a bottom plate, a sample binding pad, a nitrocellulose membrane, and a water absorbing pad, wherein the sample binding pad, the nitrocellulose membrane, and the water absorbing pad are sequentially adhered on the bottom plate in an overlapping manner, fluorescent microsphere-labeled anti-clenbuterol hydrochloride monoclonal antibodies are embedded on the binding pad, a detection zone and a quality control zone are immobilized on the nitrocellulose membrane, a clenbuterol hydrochloride hapten-carrier protein conjugate is sprayed on the detection zone, and goat anti-mouse anti-antibodies are sprayed on the quality control zone. The test paper strip of the present invention is mainly used for the clenbuterol hydrochloride residue detection, and has advantages of high sensitivity, rapid detection, easy operation, economy, practicality, and the like.

Description

technical field [0001] The invention belongs to the field of veterinary drug detection in food safety, and specifically relates to a fluorescent microsphere immunochromatographic test strip for detecting clenbuterol hydrochloride residue in a sample, a preparation method and application thereof. technical background [0002] Clebuterol is a β-receptor agonist belonging to anilines, mainly in the form of hydrochloride, which has the functions of redistributing body nutrition, inhibiting fat deposition, increasing protein content, increasing ketone body lean meat rate, Improve meat quality and promote animal growth. Therefore, driven by economic interests in recent years, a large amount of clenbuterol has been abused in animal husbandry and breeding. When the human body takes too much CL, it will cause a series of adverse reactions, mainly including muscle tremors, rapid heartbeat and breathing, etc., which will seriously endanger life. In view of this, many countries have b...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/533
CPCG01N33/533G01N33/558G01N33/582
Inventor 何方洋冯才伟罗晓琴万宇平朱亮亮杨学林冯月君龙光宗
Owner BEIJING KWINBON BIOTECH
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