Specific primer and kit for detecting common food allergens

A specific and allergen technology, applied in the PCR technology of four common food allergens of almond, soybean, celery, fish, and the sequence of PCR primers, can solve the problem of high allergenicity, and achieve high accuracy and method. The effect of simple and easy industrial production

Inactive Publication Date: 2013-04-24
TIANJIN BIOCHIP TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The eating habits of different countries and regions are different, and the adaptability of the body to food is also different, so the food allergens are also different. For example, Westerners t

Method used

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  • Specific primer and kit for detecting common food allergens
  • Specific primer and kit for detecting common food allergens
  • Specific primer and kit for detecting common food allergens

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0059] Example 1 : Genome Extraction

[0060] 1. Extraction of plant and animal tissue genomes (using TaKaRa MiniBEST Universal Genomic DNA Extraction Kit Ver.4.0 from Treasure Biological Company)

[0061] ① Take 2-30 mg of animal tissue or 25-100 mg of plant tissue, put it in a 2 ml tube, cut it into pieces as much as possible with scissors, and grind with liquid nitrogen for some hard tissues.

[0062] ② Add 180 μl of Buffer GL, 20 μl of Proteinase K and 10 μl of RNase A (10 mg / ml), warm in a 56°C water bath until the tissue is completely lysed (2 to 3 hours, and the difficult-to-lyse materials can be appropriately prolonged or even overnight. The plant material may have residual fibrous tissue that cannot be completely lysed and will not affect DNA extraction).

[0063] Note) During the warm bath, the sample can be taken out from time to time for shaking or pipetting to speed up the lysis.

[0064] ③ Add 200μl Buffer GB to the lysate and mix well by pipetting.

[006...

Embodiment 2

[0088] Embodiment 2: the design of primer

[0089] According to the sequences and references downloaded from NCBI, primers were designed for food allergen-specific genes. The primer sequences are shown in Table 1 below:

[0090] Table 1 Allergen-specific primer sequences

[0091]

Embodiment 3

[0092] Embodiment 3: Screening of specific primers

[0093] Primers were designed for food allergens celery, fish, soybean, and almond, and the primer sequences are shown in Table 1 (SEQ ID NO:1-SEQ ID NO:8). The specificity of other common food chicken, peanut, sesame and wheat was collected to verify the primers, and the source of food is shown in Table 2 below. Table 2 Foods tested

[0094] Allergen name specific gene source celery MTD Tianjin Development Zone Vegetable Market fish Mitochondrial 16S Tianjin Development Zone Vegetable Market soybean Lectin Tianjin Development Zone Vegetable Market almond Pru dul gene Tianjin Development Zone Vegetable Market chicken * Tianjin Development Zone Vegetable Market peanut * Tianjin Development Zone Vegetable Market Sesame * Tianjin Development Zone Vegetable Market wheat * Tianjin Development Zone Vegetable Market

[0095] The PCR system is ...

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Abstract

The invention relates to a specific primer and a kit for detecting common food allergens, and particularly relates to a specific PCR (Polymerase Chain Reaction) primer for detecting food allergens and a method and application thereof for rapidly detecting the common food allergens with a multiplex-PCR kit. The kit comprises a 10*PCR reaction solution, MgC12, dNTP (Diethyl-Nitrophenyl Thiophosphate), a primer and DNA (Deoxyribonucleic Acid) polymerase, wherein the primer comprises the following specific nucleotide sequences: (1) MTD (Maximum Tolerated Dose) gene of celery, (2) mitochondrial 16S of fish, (3) Lectin gene of soybean and (4) Prudul gene of almond. The specific nucleotide, the PCR kit with the nucleotide, and the gene chip or a micro-array which are used for detecting the common food allergens are high in practicability; and the PCR kit is simple and convenient to prepare, needs a short detection period, has a high speed and high accuracy, is high in operability, can reduce detection costs and is prone to industrial production.

Description

technical field [0001] The invention relates to a multiplex PCR rapid detection method and its application, in particular to a PCR technique which can be simultaneously used to detect four common food allergens of celery, fish, soybean and almond and the sequences of the PCR primers used therefor. Background technique [0002] Food allergy is also called food allergy or digestive system allergy (allergic reaction of digestive system), allergic gastroenteritis, etc. And non-IgE-mediated immune reactions, leading to allergic reactions in the digestive system or systemic. Pathogenesis: The sensitizing antigen activates the IgE plasma cells of the intestinal lamina propria to produce a large amount of IgE antibodies, which combine with mast cells and fix on the surface of these cells. When food allergens re-enter the body and combine with IgE on the surface of mast cells in the gastrointestinal mucosa, mast cells are activated and degranulated to release a series of inflammator...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
Inventor 王磊曹勃阳刘向前陈敏冯露
Owner TIANJIN BIOCHIP TECH CO LTD
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