Specific primer, kit and identification method for identifying donkey-derived components

Abstract

The invention provides a specific primer, a kit and an identification method for identifying donkey-derived components. A large number of primers are designed based on CYT B and COX-1 genes of a donkey, and are screened and researched to screen out a species-specific primer; and the primer is used for identifying the donkey-derived components in a mixture to be detected. The specific primer, the kit and the identification method provided by the invention can accurately distinguish the donkey-derived components from other adulterants, have a PCR amplification product of only 50-150bp and smallmolecular weight, ensure identification of the donkey-derived components in a large number of mixtures within a short time, and can obtain accurate, stable, reliable and highly sensitive identification results.

Description

technical field [0001] The invention belongs to the technical field of identification of traditional Chinese medicinal materials, and in particular relates to a specific primer, a kit and an identification method for identifying components derived from donkeys. Background technique [0002] Donkey-hide gelatin (ASINICORIICOLLA) is a solid gelatin made from the dry skin or fresh skin of an equine animal donkey (Equus asinm L.), which is decocted and concentrated, and can be added with an appropriate amount of rice wine, rock sugar and soybean oil. , the effect of hemostasis, for the treatment of symptoms such as blood deficiency and chlorosis, dizziness, palpitations, muscle flaccidity, restlessness and insomnia. Ejiao is a variety included in the 2015 edition of the "Chinese Pharmacopoeia". Its medicinal use has a long history. It is an important source species, especially effective in preventing diseases. There are more than 3,200 medical prescriptions about donkey-hide g...

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Problems solved by technology

Both of the above two patents are for the identification of direct products of donkeys (donkey skin or donkey meat), which is relatively low difficulty, and high-quality DNA can be obtained by direct extraction. However, donkey-hide gelatin is a deep-processed product. After long-term decoction at high temperature, the DNA is severely damaged. The DNA content in the finished product is small and the fragments are extremely short. Moreover, the main components of donkey-hide gelatin are various collagen peptides. The existence of a large amount of protein will seriously affect the efficiency of DNA extraction. In addition , in the process of preparing the finished donkey-hide gelatin, not only need to go through a long time of decocting, but also add auxiliary materials such as rice wine and soybean oil in the later stage. The addition of these auxiliary materials will increase the difficulty of extracting the DNA of the finished donkey-hide gelatin and introduce various components that inhibit the PCR reaction. The reason makes it very difficult to identify donkey-derived components in donkey-hide gelatin. When using the above-mentioned prior art methods to identify donkey-derived components in donkey-hide gelatin, problems such as failure to amplify and miscellaneous bands are prone to occur, and the accuracy, sensitivity, and specificity are poor. , are not suitable for the identification of donkey-hide gelatin

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