Quantitative detection method for donkey-derived and swine-derived components in donkey-hide gelatin liquid semi-finished product or finished product, composition and kit

Abstract

The invention relates to a quantitative detection method for donkey-derived and swine-derived components in a donkey-hide gelatin liquid semi-finished product or finished product, a composition and a kit, which belongs to the technical field of molecular biology. The method comprises: according to a donkey and swine house-keeping gene single-copy gene, designing a universal donkey and swine primer as well as a donkey and swine specificity probe, extracting a sample DNA to be tested, quantitatively detecting the donkey-derived or swine-derived components in the donkey-hide gelatin liquid semi-finished product or finished product by adopting a TaqMan qPCR method, calculating a copy number of donkey-derived and swine-derived karyogene in the unit mass of the donkey-hide gelatin liquid according to a standard curve and a Ct value of a sample to be tested, calculating a ratio of the donkey-derived components and a ratio of the swine-derived components in donkey-hide gelatin, and quantifying the donkey-derived or swine-derived components in the gelatin sample. The method, the composition and the kit are good in detection specificity, high in sensitivity, not only capable of qualitatively detecting the donkey-derived and swine-derived components in the donkey-hide gelatin, but also capable of realizing the quantitative analysis, and capable of providing technical popularization and application for the quality control of donkey-hide gelatin enterprises.

Description

technical field [0001] The present invention relates to a qualitative and quantitative detection method of donkey-hide gelatin glue and finished products, and specifically relates to a quantitative detection method, composition and kit for donkey- and pig-derived ingredients in donkey-hide gelatin glue semi-finished products or finished products, belonging to field of molecular biology. Background technique [0002] Donkey-hide gelatin is a solid gelatin made from the donkey skin of the equine animal, which is boiled and concentrated. It is originally produced in the Pandong-E region of Shandong Province. It was first recorded in "Shen Nong's Materia Medica". , has a history of nearly three thousand years. Donkey-hide gelatin blood-tonifying holy medicine, tastes sweet and flat, enters the lung, liver, and kidney meridian, has the functions of nourishing blood and stopping bleeding, nourishing yin and moistening dryness, etc. Power, applicable to a wide range of people. ...

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Problems solved by technology

[0005] In recent years, as the adulteration of donkey-hide gelatin has been continuously exposed by the media, it has gradually attracted the attention of scientific researchers. Domestic and foreign scientific research institutions and donkey-hide gelatin companies have carried out a large number of scientific research work on the identification of the authenticity of donkey hide and horse hide by means of molecular biology techniques. , the patent authorization number is (CN.104046700B) using qPCR technology to solve the technical problem of quickly identifying donkey skin, horse skin, donkey mule and horse mule skin, and the patent (CN201510018058.3) discloses the Originated multiple real-time fluorescent PCR detection method, patent (CN201510403781.3) uses nested PCR technology to identify the authenticity of donkey-hide gelatin, but all of them use mitochondria as the target gene for qualitative detection of the authenticity of donkey-hide gelatin products, because mitochondrial genes are in all animals The copy numbers in different tissue cells are different and cannot be quantified. It is impossible to distinguish whether the donkey-hide gelatin is contaminated or intentionally adulterated, and it is impossible to know the adulteration ratio. Therefore, quantitative research on the adulteration of donkey-hide gelatin is necessary

However, the quantitative analysis method of donkey and pig source components based on DNA detection technology has not yet been reported at home and abroad.

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