Method for identifying celiac disease allergen in wheat flour by high-throughput sequencing technology

Abstract

The invention relates to a method for identifying celiac disease allergen in wheat flour by a high-throughput sequencing technology, and belongs to a DNA identification technology of allergic proteincausing celiac disease allergy by taking wheat as food. The method comprises the following steps: designing three groups of primers by utilizing DNA sequences of wheat gluten causing celiac disease and prolamin, extracting genome DNA from to-be-tested wheat flour, performing PCR amplification on wheat allergen by the three groups of primers to obtain amplified DNA products; sequencing the amplified DNA products by using a high-throughput sequencing instrument to obtain gluten and prolamin DNA fragment sequences; comparing the DNA fragment sequences of two kinds of allergic protein with the DNAfragments of the various known protein short peptide celiac disease allergens by a comparison algorithm of the same segment in the two DNA sequences to obtain the quantity and the positions of the DNA fragments of the allergens. The method has the following advantages: the quantity and the positions of the DNA fragments of various kinds of wheat allergenic protein peptide can be quantitatively detected in wheat samples at one time, so the method can serve as detection technology means of a method for evaluating intensity of celiac disease caused by the wheat.

Description

technical field [0001] The invention belongs to the DNA identification technology, in particular to the DNA identification technology of the allergenic protein caused by celiac disease allergy caused by wheat as food. Background technique [0002] Celiac disease (CD), also known as wheat gluten allergy, is currently considered to be a small intestinal malabsorption syndrome caused by patients' intolerance to prolamins in wheat gluten. Symptoms are diarrhea, gastroesophageal reflux, nausea, vomiting, anemia, etc. It is a serious allergic disease caused by wheat. It is an autoimmune disease mediated by human autoantibodies IgG and IgA. Human leucocyte antigen (Human leucocyte antigen) , HLA) with DQ2 or DQ8 genotypes are susceptible populations, and the incidence rate is relatively high in North America, Northern Europe, and Australia, and it is generally missed diagnosis in China. Celiac disease is mainly induced by storage proteins of wheat crops such as wheat and barley. ...

AI Technical Summary

Problems solved by technology

[0009] Since the existing technology can only obtain whether the tested wheat contains these 17 celiac disease allergens, it is not known that a short DNA fragment of a wheat allergen protein peptide has a presence in the long DNA chain of the wheat allergen protein. How much, so in the prior art evaluation criteria for wheat allergenicity, there is no evaluation method for evaluating the strength of wheat allergenicity by using the number of DNA fragments of the same wheat allergen protein peptide

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