Quantitative detection method, composition and kit of donkey/horse-derived components in donkey-hide gelatin colloidal liquid semifinished product or finished product

Abstract

The invention relates to a quantitative detection method, composition and kit of donkey / horse-derived components in a donkey-hide gelatin colloidal liquid semifinished product or finished product, belonging to the technical field of molecular biology. The method comprises the following steps: designing donkey / horse universal primers and donkey / horse specific probes according to the donkey / horse house-keeping gene single-copy genes, extracting DNA (deoxyribonucleic acid) of a sample to be detected, carrying out TaqMan qPCR (quantitative polymerase chain reaction) on the donkey-hide gelatin colloidal liquid semifinished product or finished product to perform quantitative detection on donkey / horse-derived components, calculating the number of copies of the donkey / horse-derived nuclear genes in the unit mass of the donkey-hide gelatin colloidal liquid according to the standard curve and the Ct value of the sample to be detected, and calculating the proportion of the donkey / horse-derived components in the donkey-hide gelatin, thereby quantifying the donkey / horse-derived components in the donkey-hide gelatin sample. The method, composition and kit have the advantages of high specificity and high sensitivity, implement qualitative detection on the donkey / horse-derived components in the donkey-hide gelatin, and can also implement quantitative analysis, thereby providing technological popularization and application for quality control of donkey-hide gelatin enterprises.

Description

technical field [0001] The invention relates to a quantitative detection method, composition and kit for donkey- and horse-derived components in a semi-finished or finished product of donkey-hide gelatin glue, and belongs to the technical 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. Li Shizhen's "Compendium of Materia Medica" contains: "Ejiao's "The Classic" is top grade. [0003] The 2015 edition of the ...

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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, and it is impossible to distinguish whether the donkey-hide gelatin is contaminated or 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 horse source components based on DNA detection technology has not yet been reported at home and abroad.

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