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Gene chip and detection kit for detecting common food allergens

A technology of gene chips and allergens, applied in the field of gene chips and detection kits containing the chips, can solve problems such as shortening the detection cycle, and achieve the effects of making up for the lack of detection range, accurate detection and identification, and rapid and accurate detection

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

For this reason, in the present invention, DNA is used as the detection target, 8 kinds of food samples are selected, and a new gene chip for detecting allergens in food is established to make up for the time-consuming, labor-intensive, and poor resolution defects of traditional detection techniques. Expand the detection range, improve detection sensitivity and specificity, reduce labor intensity, and shorten the detection cycle

Method used

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  • Gene chip and detection kit for detecting common food allergens
  • Gene chip and detection kit for detecting common food allergens
  • Gene chip and detection kit for detecting common food allergens

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0078] Example 1 Probe design and preparation

[0079] 1. Sequence acquisition:

[0080] The MTD gene of celery, the mitochondrial DNA of chicken, the mitochondrial 16s gene of fish, the Arah1 gene of peanut, the Sal gene of sesame, the Lectin gene of soybean (bean sprouts), the GAG56D gene of wheat, and the Pru dul gene of almond were downloaded from the GenBank public database.

[0081] 2. Example of probe design:

[0082] (1) MTD gene probe of celery: Import the MTD gene sequence of celery into Glustal X software, select a representative sequence and perform blastn comparison in the public data NCBI to determine whether it can be used as a specific target and the position of the specific target. Import the sequence into OligoArray2.0 software. The parameters were set as follows: -n 20; -l 30; -L 40; -D 3000; -t 79; -T 90; -s 65 °C; -x 65 °C; -N 2; ; -m GGGGG CCCCC TTTTT AAAAA ; -g 15. Run the program to design probes online.

[0083] 3. Probe synthesis: Extend t...

Embodiment 2

[0091] Example 2 Primer design and preparation

[0092] 1. Example of primer design:

[0093] The MTD gene of celery, the mitochondrial DNA of chicken, the mitochondrial 16s of fish, the Arah1 gene of peanut, the Sal gene of sesame, the Lectin gene of soybean (bean sprouts), the GAG56D gene of wheat, and the Pru dul gene of almond. Import MTD gene, chicken mitochondrial DNA, fish mitochondrial 16s, peanut Arah1 gene, sesame Sal gene, soybean (bean sprouts) Lectin gene, wheat GAG56D gene, almond Pru dul gene sequence into Glustal X software, and select one of them A representative sequence was imported into the Primer Primer 5.0 software, and the length was set to 70bp~10bp, the G+C% value was 40%~60%, Hairpin: NONE, Dimer: NONE, False Priming: NONE, Cross Dimer: NONE. And look for the nucleotide sequence region suitable for primer design, its characteristics basically meet the following conditions: 1, this region should contain a variable region that is easy to design s...

Embodiment 3

[0106] Example 3 Rapid detection of common food allergens and preparation of kits using gene chips

[0107] 1. Sample handling:

[0108] (1) Buy the food needed for the experiment at the vegetable market in Tianjin Binhai Development Zone

[0109] (2) Genome was extracted according to the procedure of the genome extraction kit of Dalian Bao Biological Company.

[0110] (3) Redissolve the genome, which can be used for detection or stored at -20°C.

[0111] Attachment: Kit name: Bao Bioengineering Company DNA Extraction Kit for GMO Detection Ver.2.0)

[0112] TaKaRa MiniBEST Universal Genomic DNA Extraction Kit Ver.4.0 from Treasure Biotech Co., Ltd.

[0113] 2. Amplify the target sequence: Take 3 ul of the supernatant extracted by the above genome extraction method as a template and add it to the PCR reaction mixture. The formula of the PCR reaction mixture is shown in Table 3 below. (Note: The PCR buffer, MgCl2, dNTP mixture, and Taq enzyme in the following T...

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Abstract

The invention provides a gene chip and a detection kit for detecting common food allergens, which mainly aim at eight common food allergens of celery, chicken, fish, peanut, sesame, soybean, wheat and almond. The gene chip comprises a solid phase carrier and an oligonucleotide probe which is fixed on the solid carrier and comprises DNA (Deoxyribonucleic Acid) fragments selected from MTD (Maximum Tolerated Dose) gene of celery, mitochondrial DNA of chicken, mitochondrial 16S of fish, Arah1 gene of peanut, Sal gene of sesame, Lectin gene of soybean, GAG56D gene of wheat and Prudul gene of almond. The gene chip and the detection kit can be used for detecting the common food allergens and have the characteristics of being convenience in operation, high flux, high accuracy, high repeatability and the like, and can be used for clinical detection on food anaphylaxis.

Description

technical field [0001] The invention relates to a gene chip and a detection kit containing the chip, in particular to a common food allergen gene chip and a detection kit. Background technique [0002] Food allergy is an adverse reaction that people have to certain foods, which is an allergic reaction in medicine. According to some foreign epidemiological surveys, 4% of the total population and 6% to 7% of infants under 3 years old are allergic to certain foods. About 80% of infants and young children develop immune tolerance to milk, eggs, peanuts, wheat, and soybeans at the age of 5 to 6 years, about 20% of allergies to fish and crustaceans will be eliminated, and the rest are often lifelong allergies. In adults, the incidence of fruit and vegetable allergy is on the rise, mainly because of the fruit and vegetable-pollen immune cross-reaction. my country has not yet conducted a large-scale survey, but earlier small-scale surveys showed that in Beijing, Guangzhou and Shen...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C40B40/06
Inventor 王磊曹勃阳刘向前陈敏冯露
Owner TIANJIN BIOCHIP TECH CO LTD
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