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Fluorescent microsphere immunochromatographic test strip for detecting aflatoxin M1, and method thereof

An immunochromatographic test paper, aflatoxin technology, which is applied in the direction of testing food, testing dairy products, measuring devices, etc., can solve the problems of cumbersome operation of thin layer chromatography, low sample purity requirements, short detection time, etc. The effect of reaction sensitivity, short detection time, and simple method

Active Publication Date: 2015-09-09
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. It has the advantages of low requirements, especially suitable for the detection of large batches of samples. Compared with ELISA, the detection time of fluorescent immunochromatography is shorter, the stability is good, the operation is simple, no other equipment is needed, and the result judgment is intuitive and reliable. It is suitable for on-site Do a quick filter

Method used

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  • Fluorescent microsphere immunochromatographic test strip for detecting aflatoxin M1, and method thereof
  • Fluorescent microsphere immunochromatographic test strip for detecting aflatoxin M1, and method thereof
  • Fluorescent microsphere immunochromatographic test strip for detecting aflatoxin M1, and method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Embodiment 1 detects the composition of the fluorescent microsphere immunochromatography test strip of aflatoxin M1

[0038] The test strip is composed of a sample binding pad, a chromatographic membrane, an absorbent pad and a backing, such as figure 1 shown;

[0039] The sample binding pad 1, the chromatographic membrane 2, and the water-absorbing pad 3 are pasted on the substrate 6 in sequence, the end of the sample-binding pad is connected to the beginning of the chromatographic membrane, the end of the chromatographic membrane is connected to the water-absorbing pad, and the sample is combined. The beginning of the pad is aligned with the beginning of the backing, and the end of the absorbent pad is aligned with the end of the backing;

[0040] There are detection zone 4 and quality control zone 5 on the chromatographic membrane, both in the form of strips perpendicular to the length of the test strip, the detection zone is located near the end of the sample bindi...

Embodiment 3

[0085] The detection of aflatoxin M1 in the sample of embodiment 3

[0086] 1. Test the sample with a test strip

[0087] Take out the required number of test strips from the original package and mark them; at the same time, put the milk sample to be tested in a water bath at 60-70°C for 5 minutes, take it out; use a micropipette to draw 150 μl of the milk sample solution to be tested and add it vertically Put 3 to 4 drops (about 100 μL) in the sample hole, start timing when the liquid flows, and react for 3 to 5 minutes; insert the test strip into the carrier of the KFT-100A fluorescence detector, and select the item to be tested through the touch screen , press the "Start Test" button, the fluorescence detector will automatically scan the test card; read the test results on the display screen of the instrument.

[0088] 2. Analysis of test results

[0089] After the test is completed, the instrument will automatically calculate the concentration of aflatoxin M1 in the milk...

Embodiment 4

[0093] The determination of embodiment 4 test strip technical parameters

[0094] 1. Detection limit test

[0095] Add the aflatoxin M1 standard substance to the blank milk sample to the final concentration of 0, 0.05, 0.1, and 0.2 μg / L, and test the milk sample with a test strip. The result is: when the concentration of the aflatoxin M1 standard substance is 0 , 0.05μg / L, the result of the fluorescence detector is negative, when the concentration of aflatoxin M1 standard substance is 0.1, 0.2μg / L, the result of the fluorescence detector is positive, indicating that the test strip is effective against Aspergillus flavus in milk samples The detection limit of toxin M1 was 0.1 μg / L.

[0096] 2. False positive rate, false negative rate test

[0097] Take 20 milk positive samples with known aflatoxin M1 content greater than 0.1 μg / L and 20 milk negative samples with aflatoxin M1 content less than 0.1 μg / L, and test them with test strips produced in 3 batches , the results are s...

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Abstract

The invention discloses a fluorescent microsphere immunochromatographic test strip for detecting aflatoxin M1. The test strip comprises a sample binding pad, a chromatographic membrane, a water absorption pad and a viscose bottom liner, and the sample binding pad contains a fluorescent microsphere labeled aflatoxin M1 monoclonal antibody; and the chromatographic membrane comprises a detection zone and a quality control zone, the detection zone is coated with an aflatoxin M1 semiantigen-carrier protein conjugate, and the quality control zone is coated with a goat anti-mouse antiantibody. A method for detecting the aflatoxin M1 by using the test strip has the advantages of simplicity, rapidness, wide application range, low cost and easy popularization use.

Description

technical field [0001] The invention relates to a test strip and a method for detecting aflatoxin M1, in particular to a fluorescent microsphere immunochromatographic 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. 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, directly or indirectly enters the human food chain, and is a carcinogenic, mutagenic, and mutagenic poison. Serious threat t...

Claims

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Application Information

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IPC IPC(8): G01N33/04G01N33/577
Inventor 万宇平冯才伟赵正苗何方洋朱亮亮冯静崔海峰余厚美
Owner BEIJING KWINBON BIOTECH
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