Fluorescent quantitative PCR method and primer pair for detection of fish parvalbumin

Abstract

The invention relates to a fluorescent quantitative PCR method for detecting fish parvalbumin and a primer pair and belongs to the technical field of food safety detection. The PCR method comprises the following steps: I, designing a specific amplification primer pair; II, extracting DNA of a sample; III, judging whether the sample contains a parvalbumin gene or not through an amplification curve and a dissolution curve; the invention further relates to the primer pair. To be specific, the primer pair is as follows: base sequences of the fish parvalbumin are shown as SEQ ID No.1 and SEQ ID No.2. Compared with the prior art, the fluorescent quantitative PCR method and the primer pair have the advantages as follows: by using bioinformatics and comparative genomics, the shared gene sequences of fish parvalbumin are selected, the specific amplification primer pair is designed according to the sequences, the primer pair provided by the invention can be used for carrying out fluorescent quantitative PCR detection on samples to detected and can rapidly, sensitively and specifically detect the allergen ingredient namely parvalbumin in different kinds of fishes and fish products in food.

Description

technical field [0001] The invention relates to the technical field of food safety detection, in particular to a fluorescent quantitative PCR method and a primer pair for detecting fish parvalbumin. Background technique [0002] In 1969, Aas and Elsayed identified the main allergen of Pacific cod (Gadus callarias) for the first time and named it Gad c 1, which was one of the first identified food allergens in the world. Gad c 1 is composed of 113 amino acids and 1 glucose molecule, with a molecular weight of about 12kD. It belongs to parvalbumin and is a calcium-binding protein. This parvalbumin exists in large quantities in the white muscles of lower vertebrates. Parvalbumin can be divided into two different types according to the difference in amino acid sequence: α-type parvalbumin pI≥5.0, containing a small amount of acidic parvalbumin, and β-type parvalbumin pI≤4.5, with higher acidic parvalbumin content. Most of the parvalbumin in fish belong to the β type. [0003] ...

AI Technical Summary

Problems solved by technology

The current detection methods for fish parvalbumin mainly include serological testing and protein characteristic methods, among which, serological methods (RAST, CAP, etc.) The allergen solid-phase carrier has different binding ability to IgE, so it is more difficult to commercialize food allergen detection; and the ELISA analysis based on protein characteristics is based on animal antibodies with stable quality and specificity. Good sensitivity and sensitivity, especially for deep-processed products, the stability of heat-stable proteins is higher than that of DNA, but it is difficult to obtain a highly specific antibody

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