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Colloidal gold immunochromatographic test strip for detecting apricot kernel allergen in food and preparation method thereof

A technology of immunochromatography test paper and colloidal gold, which is applied in the direction of analysis materials, measuring devices, instruments, etc., can solve the problems of long detection time and high cost, and achieve the effect of high sensitivity and strong specificity

Inactive Publication Date: 2017-11-07
ANHUI UNIVERSITY +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, almond allergy detection methods include ELISA, immunosensor detection technology, real-time fluorescent quantitative PCR technology, SPR, mass spectrometry technology, etc. These detection methods have high requirements for equipment, surface environment, operators, etc., and the detection time is long and the cost is high. High, so it is necessary to establish a new type of rapid and stable test strip detection

Method used

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  • Colloidal gold immunochromatographic test strip for detecting apricot kernel allergen in food and preparation method thereof
  • Colloidal gold immunochromatographic test strip for detecting apricot kernel allergen in food and preparation method thereof
  • Colloidal gold immunochromatographic test strip for detecting apricot kernel allergen in food and preparation method thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0029] Embodiment 1, the preparation of the colloidal gold immunochromatography test strip of almond allergen in food

[0030] The preparation method of the colloidal gold immunochromatographic test strip of almond allergen in food comprises the following steps:

[0031] (1) Solution preparation:

[0032] Preparation of trisodium citrate: Accurately weigh 0.1g of trisodium citrate, dissolve it in ultrapure water, and dilute to 10mL, filter through a 0.22μm filter membrane, and prepare and use immediately;

[0033] Preparation of chloroauric acid solution: Accurately weigh 1.0g of trisodium citrate, dissolve it in ultrapure water, and dilute to 100mL, filter with a 0.22μm filter membrane, and store the brown reagent bottle at 4°C for later use;

[0034] Preparation of potassium carbonate solution: Accurately weigh 1.38g of potassium carbonate, dissolve it in ultrapure water, and dilute to 50mL, filter through a 0.22μm filter membrane, and store at 4°C for later use;

[0035] ...

Embodiment 2

[0050] Embodiment 2, the sensitivity determination of the colloidal gold immunochromatography test strip of almond allergen in food

[0051] Use the sample diluent to dilute the standard to different concentrations: 100000ppm, 10000ppm, 100ppm, 10ppm, 5ppm, 2ppm, 1ppm, use the test paper prepared in Example 1 for detection, use the sample diluent as a negative control (0ppm), observe after 5min As a result, the detection results are as figure 1 shown. The results show that 100,000ppm is positive at a high concentration, and the C line is colored; 100-10,000ppm is positive at a higher concentration, and the TC line is colored; 2-10ppm is positive, and the THC line is colored; 0ppm is a negative control, and the HC line is colored. The test strip has a sensitivity of 2 ppm.

Embodiment 3

[0052] Embodiment 3, the specificity determination of the colloidal gold immunochromatography test strip of almond allergen in food

[0053] Get respectively peanut, wheat, hazelnut as test sample, after grinding and crushing, dilute to 100ppm, detect with the test strip prepared in embodiment 1, observe the result after 5min, the test result is as follows: figure 2 shown. The results showed that peanuts, wheat, and hazelnuts all showed C lines at 100ppm, and the test strip had no cross-reaction with peanuts, wheat, and hazelnuts, showing that the test strip had good specificity.

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Abstract

The invention discloses a colloidal gold immunochromatographic test strip for almond allergens in food and a preparation method thereof. The chromatographic test strip comprises a water-absorbing pad, a nitrocellulose membrane, a gold standard pad and a sample pad assembled in sequence, and also includes Loading bottom plate; detection line, antigen line and quality control line are set on the nitrocellulose membrane, the gold label pad contains a colloidal gold-labeled anti-almond protein monoclonal antibody, and the detection line of the nitrocellulose membrane is coated with another Capture the monoclonal antibody against almond protein, the antigen line is coated with almond protein, and the quality control line is coated with goat anti-mouse IgG; when a quality control line and an antigen line are negative on the test strip, a test line appears (regardless of whether there is antigen or not) line) are all positive, compared with the existing almond protein allergen detection technology, it has high sensitivity and strong specificity, and can also make an intuitive evaluation of the concentration of the allergen, which can be directly used for food detection.

Description

technical field [0001] The invention belongs to the technical field of food safety detection, and in particular relates to a colloidal gold immunochromatographic test strip for detecting almond allergens in food and a preparation method thereof. Background technique [0002] Almonds, taxonomically called Amygdala, belong to the family Amylaceae, and are nut allergens. Almonds are present in many processed foods, which may pose a potential allergenic hazard. According to related reports, the main storage protein of almonds is amandin, which contains the main reaction polymorphisms recognized by the patient's serum IgE, and is the main allergen that causes illness in patients with almond allergy. At present, almond allergy detection methods include ELISA, immunosensor detection technology, real-time fluorescent quantitative PCR technology, SPR, mass spectrometry technology, etc. These detection methods have high requirements for equipment, surface environment, operators, etc....

Claims

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Application Information

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IPC IPC(8): G01N33/558G01N33/577
CPCG01N33/558G01N33/577
Inventor 唐恒立周祚琴赵亮杰森 M.罗博特姆肯尼斯 H.鲁亨利 格赖斯张部昌唐小迪
Owner ANHUI UNIVERSITY
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