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Primer for detecting allergen gene in scallop

An allergen and gene technology, applied in the field of primer sequences used for rapid detection of scallop allergen gene tropomyosin

Inactive Publication Date: 2015-01-07
INSPECTION & QUARANTINE TECH CENT SHANDONG ENTRY EXIT INSPECTION & QUARANTINE BUREAU +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The reaction can accumulate target DNA to 10 within one hour. 9 ~10 10 copy

Method used

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  • Primer for detecting allergen gene in scallop
  • Primer for detecting allergen gene in scallop

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Follow the procedure below for testing:

[0025] (1) Extraction of DNA from shellfish samples to be tested

[0026] A. Weigh 10g sample, grind it into powder with liquid nitrogen, transfer 0.1g to a 1.5mL centrifuge tube. Add 600 μL of preheated lysis buffer and 20 μL of proteinase K, mix gently, and incubate in a water bath at 65°C for 3 hours;

[0027] B. Add an equal volume of 600 μL of phenol / chloroform into the tube, mix well by inverting up and down, and extract for 2 minutes;

[0028] C. Centrifuge at 12000g for 5min, draw the supernatant into a new centrifuge tube, add the same volume of sedimentation solution as the supernatant, mix well, leave at room temperature for 10min, centrifuge at 12000g for 5min, remove the supernatant, and keep the precipitate;

[0029] D. Add 60 μL of RNase to the precipitate, place it at 37°C for 2 minutes, mix it well with a pipette tip, dissolve the precipitate at 37°C, add 300 μL of buffer solution after 5 minutes, and mix up a...

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PUM

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Abstract

The invention belongs to quick screening and detection techniques for allergen tropomyosin gene of scallop, and in particular relates to a primer group for detecting allergen gene tropomyosin of shellfish. In the invention, the specific primers are designed according to the conserved region of the tropomyosin gene. By using the primer pair and adopting the dye real-time fluorescence PCR (polymerase chain reaction) amplification technology, the tropomyosin gene in the scallop can be quickly, sensitively and specifically detected, thereby screening whether the product contains allergen components of scallop. The primers can also be provided in a kit form together with other reagents for nucleic acid amplification reactions, and the method is easy and simple to operate and has good repeatability.

Description

technical field [0001] The invention relates to a method for rapid screening and detection of allergen genes in scallops by using dye method real-time fluorescent polymerase chain reaction technology, in particular to a primer sequence used for rapid detection of scallop allergen gene tropomyosin. Background technique [0002] Shellfish is one of the highest-quality edible protein resources for human beings, and it is also one of the eight types of allergic foods announced by the Food and Agriculture Organization of the United Nations. Shellfish allergies seriously affect the health and quality of life of allergic people. Detection research is very necessary. For people with allergies, a small amount of ingestion or even contact with shellfish protein may lead to allergies. Therefore, reliable shellfish allergen detection methods will help standardize aquatic food labels and ensure food safety for consumers. Although patients with shellfish allergies can avoid allergens acc...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68
Inventor 刘彩霞孙敏林超高宏伟梁成珠徐彪
Owner INSPECTION & QUARANTINE TECH CENT SHANDONG ENTRY EXIT INSPECTION & QUARANTINE BUREAU