Quantitative determination method for donkey and bovine derived ingredients in colla corii asini liquid semi-finished product or finished product, composition and kit

Abstract

The invention relates to a quantitative determination method for donkey and bovine derived ingredients in a colla corii asini liquid semi-finished product or finished product, a composition and a kit, and belongs to the technical field of molecular biology. According to the method, donkey and bovine general primers and donkey and bovine specific probes are designed according to donkey and bovine house-keeping gene single-copy genes; DNA (Deoxyribonucleic Acid) of a sample to be determined is extracted; the colla corii asini liquid semi-finished product or finished product is subjected to donkey or bovine derived ingredient quantitative determination by a TaqMan qPCR method; the copy number of donkey and bovine derived nuclear genes in the unit mass of the colla corii asini liquid is calculated according to a standard curve and the Ct value of the sample to be determined; the proportions of the donkey and bovine derived ingredients are calculated; and further, the donkey and bovine derived ingredients of the colla corii asini sample are further quantified. The method, the composition and the kit have the advantages that the detection specificity is good; the sensitivity is high; the donkey and bovine derived qualitative determination in the colla corii asini can be realized; the quantitative analysis can also be realized; the quality control can be achieved by colla corii asini enterprises; and the technical popularization and application can be realized.

Description

technical field [0001] The present invention relates to a quantitative detection method of donkey-hide gelatin glue and animal-derived components in finished products, in particular to a quantitative detection method, composition and kit for donkey- and bovine-derived ingredients in donkey-hide gelatin glue semi-finished products or finished products, belonging to molecular biology field of technology. 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 ran...

AI Technical Summary

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 Genetic multiple real-time fluorescent PCR detection method, patent (CN 201510403781.3) 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 different in all animals The copy numbers in tissue cells are different and cannot be quantified. It is impossible to distinguish whether the donkey-hide gelatin is contaminated or intentionally adulterated, let alone know the adulteration ratio. Therefore, quantitative research on the adulteration of donkey-hide gelatin is necessary

However, based on DNA detection technology, the quantitative analysis method of donkey-derived and bovine-derived components in donkey-hide gelatin has not yet been reported at home and abroad.

Images