Milk immunofluorescence detection test strip and application

A technology of immunofluorescence detection and test strips, which is applied in the direction of testing food, testing dairy products, measuring devices, etc., can solve the problem that the allergen labeling method does not give specific regulations, consumers' allergic reactions have not attracted enough attention, and the results not quite accurate

A technology of immunofluorescence detection and test strips, which is applied in the direction of testing food, testing dairy products, measuring devices, etc., can solve the problem that the allergen labeling method does not give specific regulations, consumers' allergic reactions have not attracted enough attention, and the results not quite accurate

CN112362865APending Publication Date: 2021-02-12杭州福德敏生物技术有限公司
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  • Milk immunofluorescence detection test strip and application
  • Milk immunofluorescence detection test strip and application

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preparation example Construction

[0023] The preparation method of the conjugate pad of the present invention preferably comprises:

[0024] (a) Adsorbing and combining fluorescently labeled streptavidin and latex microspheres to prepare fluorescent latex microspheres;

[0025] (b) combining biotin with the mixed antibody to prepare a biotinylated mixed antibody;

[0026] (c) mixing the fluorescent latex microspheres with the biotinylated mixed antibody to obtain a fluorescent latex microsphere-labeled mixed antibody;

[0027] (d) Spraying the mixed antibody labeled with fluorescent latex microspheres on the binding pad; there is no time sequence between steps (a) and (b).

[0028] In the present invention, the mass ratio of fluorescently labeled streptavidin and latex microspheres described in step (a) is preferably 1:40; the volume ratio of biotin and mixed antibody described in step (b) is preferably 1:4; The volume ratio of fluorescent latex microspheres and biotinylated mixed antibodies in step (c) is p...

Embodiment 1

[0037] 1. Preparation of binding pads

[0038] 1) Preparation of fluorescent latex microspheres: Dilute latex microspheres with a particle size of 400 nm to a final concentration of 30 mg / ml with an adsorption buffer (50 mM, citrate buffer at pH 5.8) and a volume of 6 ml to obtain latex microspheres Suspension; add red fluorescein rhodamine-labeled liantavidin in the adsorption buffer according to the volume ratio of 1:50~500, the final volume is 6ml; The mixed solution was prepared in the adsorption buffer solution of mycoavidin labeled clearly; the resulting mixed solution was incubated at room temperature for 1 to 2 hours, and stirred continuously, and then centrifuged to collect the precipitate, and the precipitate was stored in buffer solution (containing 0.06% BSA Adsorption buffer) was dissolved, stored at 4°C, and set aside.

[0039] 2) Preparation of biotinylated mixed antibody

[0040] Anti-casein antibody (ab166596 of Abcam Company, GTX37769 of Gene Tex Company); ...

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Abstract

The invention provides a milk immunofluorescence detection test strip and application, and relates to the technical field of detection test strips. The immunofluorescence detection test strip comprises a sample pad, a conjugate pad, a nitrocellulose membrane and absorbent paper which are sequentially assembled and adhered to a PVC bottom plate; the conjugate pad is coated with a fluorescent latexmicrosphere labeled mixed antibody, the nitrocellulose membrane is sequentially coated with an anti-casein antibody (T1 line), an anti-beta lactoglobulin antibody (T2 line), an anti-alpha whey proteinantibody (T3 line), an anti-lactoferrin / bovine serum albumin antibody (T4 line) and a rabbit anti-mouse IgG antibody (C line), and the T1 line, T2 line, T3 line and T4 line are quality control lines.The method can be used for accurately and quantitatively detecting the contents of casein, anti-beta lactoglobulin, anti-alpha whey protein and anti-lactoferrin / bovine serum albumin in food, and is simple and convenient to operate, high in accuracy and high in sensitivity.

Description

technical field [0001] The invention belongs to the technical field of detection test strips, in particular to a milk immunofluorescence detection test strip and its application. Background technique [0002] Food allergens refer to the antigenic substances present in food that can selectively activate human cells, induce specific antibody responses, and cause allergic reactions. Generally speaking, food allergens are proteins or glycoproteins with a molecular weight between 10,000 and 70,000. Common food allergens include milk, beans, eggs, cereals, tree fruits, shellfish, etc. According to the World Health Organization report, among the factors that threaten human health, food allergy ranks No. 4. In recent years, among domestic and foreign food-related warning notifications, the number of allergen issues ranks second, second only to microbial contamination, and has become a hot spot in the industry that is widely concerned internationally. The issue of food allergens h...

Claims

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Application Information

Patent Timeline
12 Feb 2021
Publication
CN112362865A
IPC
G01N33/533; G01N33/543; G01N33/558
CPC
G01N33/54313; G01N33/558; G01N33/533; G01N33/04; G01N33/54346; G01N33/54386; G01N2800/24; G01N33/54388
Inventors
武涌; 李欣