A rapid detection method and kit for donkey-derived components in food

Abstract

The invention provides a rapid detection kit for donkey source components in food, and relates to the technical field of biological species identification. The kit involves the steps that firstly a donkey source common internal standard gene LOC106825524 is screened out, the nucleotide sequence of the gene is shown in SEQ ID NO.1, the copy number of the gene is constant in a donkey species, and the gene does not have allelic variation and can serve as a target gene for identifying a donkey source. An LAMP primer is designed by taking the gene as a target sequence, and is subjected to isothermal amplification with an LAMP reaction liquid together, and an LAMP reaction product is used for colloidal gold nucleic acid test strip detection. An LAMP reaction combines with the colloidal gold nucleic acid test strip detection to form the rapid detection kit, so that the donkey source components in the food can be detected rapidly and sensitively, and the detection sensitivity can reach 0.2% (w / w). The kit has the advantages that the use method is simple, the cost is low, the reaction is easy to observe, the specificity is good, and the kit is very suitable for real-time detection on site.

Description

technical field [0001] The invention relates to the technical field of identification of biological species, in particular to a rapid detection kit for detection of donkey-derived components in foods using a loop-mediated isothermal amplification technique (LAMP) combined with colloidal gold nucleic acid test strip detection. Background technique [0002] With the rapid development of the economy and the improvement of people's living standards, the demand of Chinese residents for meat products is increasing year by year. Although many countries clearly require food labels to clearly indicate the type and source of meat and prohibit adulteration, there are still many incidents of meat adulteration in the market. For example, in order to reduce costs, donkey meat is mixed with pork Or horse meat, beef mixed with pork, sausage mixed with other meats and so on. At present, the detection method for food adulteration is mainly PCR method. The PCR method needs to rely on special...

AI Technical Summary

Problems solved by technology

At present, the adulteration detection technology of meat products at home and abroad is mostly based on the design of specific primers for the genes on the mitochondria to perform real-time fluorescent quantitative PCR amplification. Since the mitochondrial genes are multi-copy genes, their detection sensitivity is high, but at the same time it is difficult to distinguish due to sales, transportation Confusion between unintentional cross-contamination and intentional illegal additions

In addition, the concentration of high-copy mitochondrial DNA cannot correspond to the concentration of genomic DNA, so it is impossible to perform accurate quantitative analysis on samples. At the same time, due to the high homology of mitochondrial genes, it is difficult to achieve qualitative detection by ordinary PCR

Therefore, this method can only realize the screening and identification of meat adulteration by means of fluorescent quantitative PCR

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