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Time-resolved fluorescent immunochromatographic test strip for detecting tetracyclines and preparation method and application thereof

A tetracycline-based drug, time-resolved fluorescence technology, applied in analytical materials, biological tests, measurement devices, etc., can solve the problem of inappropriate large-throughput rapid screening detection, high professional requirements for operators, and difficult to achieve multiple detection and detection. Joint inspection and other problems, to achieve the effect of reducing the background fluorescence intensity, increasing the reaction sensitivity, and reducing the interference of specific stray light on the detection

Inactive Publication Date: 2018-09-07
BEIJING KWINBON BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, since the above methods all require advanced detection instruments, the detection costs are expensive, the steps are cumbersome and time-consuming, and the requirements for the professionalism of operators are high, 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 the detection of tetracyclines at home and abroad, but there are still some defects, such as poor stability, low sensitivity, and unrealistic Quantitative detection, matrix effect is obvious, 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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  • Time-resolved fluorescent immunochromatographic test strip for detecting tetracyclines and preparation method and application thereof
  • Time-resolved fluorescent immunochromatographic test strip for detecting tetracyclines and preparation method and application thereof
  • Time-resolved fluorescent immunochromatographic test strip for detecting tetracyclines and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Embodiment 1 detects the composition of the time-resolved fluorescent immunochromatography test strip of tetracyclines

[0039] 1. Test paper ( figure 1 , figure 2 )

[0040] The test paper comprises a bottom plate and a sample absorbent pad, a nitrocellulose membrane and a water absorbent pad successively lapped and pasted on the bottom plate;

[0041] The sample absorbent pad 1, nitrocellulose membrane 2 and water absorbent pad 3 are overlapped and pasted on the bottom plate 6 in sequence, the end of the sample absorbent pad is connected to the beginning of the nitrocellulose membrane, and the end of the nitrocellulose membrane is connected to the water absorbent pad. The start of the sample absorbent 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;

[0042] A detection area 4 and a quality control area 5 are fixed on the nitrocellulose membrane, and the detection area is sprayed with ...

Embodiment 2

[0047] Example 2 Preparation of Time-Resolved Fluorescence Immunochromatography Test Strips for Detecting Tetracyclines

[0048] The preparation method of the time-resolved fluorescent immunochromatographic test strip for detecting tetracycline drugs mainly includes the following steps:

[0049] 1) Preparation of microwell reagents: use fluorescent microspheres to label monoclonal antibodies to tetracycline drugs, dilute them with a specific buffer system, place them in microwells, and vacuum freeze-dry them for later use;

[0050] 2) Preparation of nitrocellulose membrane: Spray the tetracycline drug hapten-carrier protein conjugate onto the detection area range on the nitrocellulose membrane to make the detection area; spray goat anti-mouse anti-antibody onto the nitrocellulose membrane The scope of the quality control area above is made into the quality control area;

[0051] 3) Assembling and cutting: Paste the sample absorbent pad, the nitrocellulose membrane fixed with ...

Embodiment 3

[0094] Example 3 Application of Time-Resolved Fluorescence Immunochromatography Test Strips for Detecting Tetracyclines

[0095] 1. Sample pretreatment

[0096] Accurately weigh 1g±0.01g of tissue samples into a 10mL centrifuge tube, add 3mL of sample extract, vortex for 2min to mix thoroughly; centrifuge at 3000r / min for 5min, and take the supernatant (if there is floating material in the upper layer, remove the supernatant) pending inspection.

[0097] 2. Test with test strips

[0098] Accurately draw 100 μL of the sample solution to be tested into the microwell with a micropipette, carefully pump until it is fully mixed with the reagent in the microwell, and act at room temperature (20-25°C) for 3 minutes; absorb the entire amount of the mixed solution in the well (about 100 μL ) vertically drop into the sample hole (S hole) of the test paper card, start timing when the liquid flows, and react at room temperature for 5 minutes; insert the test paper card into the carrier of...

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Abstract

The invention discloses a time-resolved fluorescent immunochromatographic test strip for detecting tetracyclines and a preparation method and application thereof. The test strip comprises test paper and a microporous reagent, the test paper comprises a bottom plate and a sample absorbing pad, a nitrocellulose membrane and a water absorbing pad which are sequentially overlapped and bonded to the bottom plate, a detecting area and a quality control area are arranged on the nitrocellulose membrane, hapten-carrier protein conjugate of the tetracyclines is sprayed on the detecting area, goat anti-mouse antiantibody is sprayed on the quality control area, and the monoclonal antibody of the tetracyclines marked by fluorescent microspheres is frozen dry in the microporous reagent. The invention further provides the preparation method of the test strip and a method for detecting the tetracyclines in samples by using the test strip. The test strip and the detecting method have the advantages ofbeing simple in operation, high in sensitivity, fast in detecting speed and low in cost, and the rapid detecting and on-spot monitoring of the tetracyclines in samples in large batches can be achieved.

Description

technical field [0001] The invention belongs to the field of detection of illegal additives in food safety, and in particular relates to a time-resolved fluorescent immunochromatographic test strip for detecting tetracycline drugs, a preparation method and application thereof. technical background [0002] Tetracyclines (TCs) have broad-spectrum and high-efficiency bactericidal and antimicrobial effects, and can be mixed with most drugs or feed additives, and have been widely used in livestock and poultry production. However, excessive use inevitably leads to serious drug residues, which has become one of the food safety hazards that seriously endanger human health. Therefore, countries all over the world pay great attention to the detection of TCs residues. my country has stipulated that the residual limit of oxytetracycline, tetracycline and chlortetracycline in milk is 100 μg / L; EU No. 675 / 92 preliminarily stipulates that the total amount of tetracycline compounds in musc...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/577
CPCG01N33/577G01N33/68
Inventor 何方洋万宇平李旭宋灏吴小胜王琳琛冯才伟王兆芹韩光耀周苑
Owner BEIJING KWINBON BIOTECH
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