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Visible chip-based method for detecting food allergens

A detection method and allergen technology, which is applied in the field of food allergen detection based on visual chips, can solve the problems of not covering food allergens, complex components, and missed detection of detection kits, so as to save time, the method is simple and fast, cost reduction effect

Inactive Publication Date: 2013-03-27
CHINESE ACAD OF INSPECTION & QUARANTINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the current food allergen kits do not cover all food allergens, and some food allergen detection kits have defects such as missed detection; in addition, the current kit detection is usually only for one allergen, but the actual In general, food ingredients are complex and there are often multiple allergens, so it is very necessary to develop high-throughput detection products to achieve one-time detection of multiple allergens

Method used

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  • Visible chip-based method for detecting food allergens
  • Visible chip-based method for detecting food allergens
  • Visible chip-based method for detecting food allergens

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Eight kinds of foods including soybean, wheat, cashew nut, peanut, milk, egg, fish and shrimp were selected as test samples.

[0047] According to the above-mentioned steps, the DNA of the above-mentioned 8 kinds of foods was subjected to multiplex PCR amplification with the primers shown in Table 1, and the multiplex PCR amplification products were hybridized with the visible film sensor chip prepared according to the above-mentioned steps, and the Bacterial water was used for blank control experiment, and the reaction results were as follows: figure 2 shown.

[0048] The signal point with the same brightness as the positive control was used as the specific reaction mark, and the signal photo was visualized through the hybridization results of the 8 food allergens ( figure 2 ) It can be seen that the positive control points are arranged in a cross-shaped pattern according to the experimental design, and the target probes all present obvious signals, which can be cle...

Embodiment 2

[0050] Take 9 pieces of visible thin-film sensor chips ordered according to the above method, use single DNA of cashew nuts and shrimp, mixed DNA of soybeans and cashew nuts, mixed DNA of milk and shrimp, mixed DNA of soybeans, cashew nuts and wheat, and mixed DNA of milk, shrimp and fish DNA, mixed DNA of milk, shrimp, fish and eggs, mixed DNA of soybean, wheat, peanut and cashew nuts as templates, multiplex PCR amplification with 8 pairs of primers shown in Table 1 as primers, and chip hybridization of PCR amplification products Experiment, and blank control experiment with sterilized water, the results are as follows image 3 shown.

[0051] It can be seen from the visual signal photos of the hybridization results that the positive control points are arranged in a cross-shaped pattern according to the experimental design, and the target probes also show obvious signals, which can be clearly distinguished from the background. Under the double assurance of PCR product specif...

Embodiment 3

[0053] Utilize the multiplex PCR method as described in embodiment 2 to buy 5 kinds of foods in the supermarket (Yana white chocolate strawberry granola bar, Orion crispy chocolate, rooster straight spaghetti, Swiss triangle dark chocolate and tempeh dace) Perform amplification, hybridize the multiple PCR amplification products with the chip, and conduct a blank control experiment with sterilized water to verify the accuracy of the food label.

[0054] Among the 5 types of food, 4 types contained milk components, 4 types contained wheat components, 3 types contained soybean components, 1 type each contained fish and egg components, and 1 type contained trace amounts of nuts.

[0055] A variety of PCR amplification products were hybridized with the chip, and sterilized water was used instead of template DNA as a blank control. Experimental results such as Figure 4 As shown, the results show that the target probes all present obvious signals, which can be clearly distinguished...

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Abstract

The invention discloses a method for detecting a plurality of food allergens in a sample, which comprises: (1) preparing a visible thin film sensor chip, wherein a plurality of specific probes corresponding to the DNA sequences of the plurality of allergens are fixed on the surface of a chip substrate respectively; (2) performing multiplex polymerase chain reaction (PCR) amplification on the DNA sequence of a test sample by using the plurality of pairs of specific upstream and downstream primers corresponding to the DNA sequences of the plurality of allergens, wherein the 5' terminal of the downstream primer has biotin bonded by a covalent bond; (3) crossing the product of the PCR amplification in the step (2) with the probes on the visible chip, and washing; (4) crossing horseradish peroxidase-marked biotin with the chip, and eluting uncrossed chains; (5) processing and developing the chip by using a horseradish peroxidase substrate; and (6), determining if the sample has the allergens and the types of the allergens according to the generation condition of a visible signal and the position of the probe corresponding to the generated visible signal. The method can quickly and synchronously detect if the sample has various allergens with high throughput.

Description

technical field [0001] The present invention generally relates to a detection method based on a visual chip, in particular to a detection method of food allergens. Background technique [0002] Food allergy is a kind of adverse reaction that people produce to food, belongs to a kind of allergic reaction that body produces to exogenous substance (allergen). Clinical manifestations of food allergy include clinical symptoms such as headache, asthma, allergic rhinitis, urticaria, enteric syndrome, anaphylactic shock, or death. Food allergy is also a food safety issue that has attracted increasing attention in recent years. Currently, the eight major allergens (foods) identified by the World Health Organization (WHO) are wheat, peanuts, soybeans, nuts, milk, eggs, fish and crustaceans. At present, there is no specific method for the treatment of food allergies. Strictly avoiding foods containing allergic ingredients is the most effective therapy. For this reason, the United St...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68
Inventor 陈颖王玮吴亚君杨海荣袁飞
Owner CHINESE ACAD OF INSPECTION & QUARANTINE
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