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Test strip and method for detecting aflatoxin M1

A technology of aflatoxin and test strips, which is applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of cumbersome operation of thin-layer chromatography, low sample purity requirements, unsuitable for rapid detection, etc., and increase reaction sensitivity , Short detection time, easy to popularize and use

Active Publication Date: 2014-01-15
BEIJING KWINBON BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Thin-layer chromatography is cumbersome to operate, heavy in pollution, poor in quantification, and time-consuming; high-performance liquid chromatography has the characteristics of high sensitivity, strong separation ability, good specificity, and reliable determination results. Before separation, the sample needs to be thoroughly and effectively purified, which is not suitable for the detection of large-scale samples, and its application is limited; enzyme-linked immunosorbent assay is a commonly used detection method at present. The advantages of low requirements are especially suitable for the detection of large batches of samples, but it is not suitable for rapid on-site detection due to the need for microplate readers and skilled operators; colloidal gold immunochromatography has a shorter detection time and is more stable than ELISA. Good performance, easy operation, no need for other instruments and equipment, intuitive and reliable judgment of results, suitable for rapid screening on site, and of great significance for the control of aflatoxin pollution

Method used

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  • Test strip and method for detecting aflatoxin M1
  • Test strip and method for detecting aflatoxin M1
  • Test strip and method for detecting aflatoxin M1

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Embodiment 1 Detects the constitution of the test strip of aflatoxin M1

[0043] 1. Test paper ( figure 1 )

[0044] The test paper is composed of a bottom plate, a sample absorption pad, a reaction film, a water absorption pad, and a protective film;

[0045] The sample absorbent pad 1, the reaction film 2, the water absorbent pad 3 and the protective film 7 are pasted on the bottom plate 6 in sequence, the end of the sample absorbent pad is connected with the reaction film, the end of the reaction film is connected with the water absorbent pad, and the sample absorbent pad The beginning is aligned with the beginning of the bottom plate, and the end of the absorbent pad is aligned with the end of the bottom plate;

[0046] The end of the sample absorption pad of the test paper is pasted with a protective film, and the protective film 7 covers the detection end on the sample absorption pad, and the word MAX is printed on the detection end protective film ( figure 2...

Embodiment 2

[0052] The preparation method of test strip described in embodiment 2 embodiment 1

[0053] 1. Preparation of test strips

[0054] The preparation method of test strip mainly comprises the following steps:

[0055] 1) Prepare freeze-dried microporous reagents with aflatoxin M1 monoclonal antibody-colloidal gold markers;

[0056] 2) Prepare a reaction membrane with a detection area coated with aflatoxin M1 hapten-carrier protein conjugate and a quality control area coated with goat anti-mouse anti-antibody;

[0057] 3) Assemble the reaction film prepared in 2) with the sample absorbent pad, water absorbent pad, protective film, and bottom plate to form a test paper;

[0058] 4) Assemble the freeze-dried microporous reagent and test paper prepared in 1) and 3) with aflatoxin M1 monoclonal antibody-colloidal gold marker into a test strip.

[0059] The following step-by-step detailed description:

[0060] (1) Preparation of each component

[0061] 1. Synthesis and identificat...

Embodiment 3

[0102] The detection of aflatoxin M1 in the sample of embodiment 3

[0103] 1. Test samples with test strips

[0104] Take out the required number of microwell reagents and test papers from the original package, and mark them; draw the milk sample to be tested, pipette 200 μl into the microwell with a micropipette, slowly suck and fully mix with the reagent in the microwell Evenly, incubate at room temperature (20°C-25°C) for 5 minutes; insert the end of the printed "MAX" line down into the microwell to fully immerse it in the solution; after incubating at room temperature (20°C-25°C) for 5 minutes, take out the test paper, judgement result.

[0105] 2. Analysis of test results

[0106] Positive: When the quality control area (C) shows a band and the detection area (T) does not develop color, it is judged as positive, and it is indicated by "+", such as Figure 4 a; Negative: When both the quality control area (C) and the detection area (T) show bands, it is judged as negati...

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Abstract

The invention discloses a test strip and a method for detecting aflatoxin M1. The test strip comprises test paper and a micro-pore reagent, wherein a colloidal gold marked aflatoxin M1 monoclonal antibody is formed in the micro-pore reagent through freeze-drying; the test paper is composed of a sample absorption pad, a reaction film, a water absorption pad, a protection film and a bottom plate, which are sequentially connected; the reaction film comprises a detection area and a quality control area, wherein the detection area is coated with an aflatoxin M1 semi-antigen-carrier protein conjugate, and the quality control area is coated with an anti-antibody. The method for detecting the aflatoxin M1 by the test strip disclosed by the invention is simple, fast, direct and accurate; furthermore, the test strip has wide application scope and low cast and is suitable for popularizing and using.

Description

technical field [0001] The invention relates to a test strip and a method for detecting aflatoxin M1, in particular to a colloidal gold test strip for detecting aflatoxin M1 in milk. Background technique [0002] Aflatoxin (AF) is the strongest type of biological toxin found to pollute agricultural products so far, and it is also a strong carcinogen. It is a class of metabolites produced by various fungi such as Aspergillus flavus and A. parasiticus, a group of compounds with similar structures. The most common ones are AF B1, B2, G1, and G2, among which AF B1 has the largest amount and toxicity. Aflatoxin M1 is the 4-hydroxyl derivative of aflatoxin B1. It is a metabolite derived from the hydroxylation of B1 in vivo. Part of it is excreted in urine and milk, and the other part exists in the edible parts of animals. It mainly exists in moldy products such as soil, animals and plants, moldy grains, various nuts, food, condiments, cooking oil, milk and milk products, directl...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N33/558G01N33/532
CPCG01N33/577G01N2333/38
Inventor 万宇平何方洋罗晓琴崔海峰冯静刘玉梅余厚美赵正苗
Owner BEIJING KWINBON BIOTECH
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