Method for identifying traditional Chinese medical material corn cervi pantotrichum based on PCR-RFLP

Abstract

The invention discloses a method for identifying the traditional Chinese medical material corn cervi pantotrichum based on PCR-RFLP. A reagent kit used for identifying the traditional Chinese medical material corn cervi pantotrichum comprises a primer pair shown in (a) or (b) and restriction enzyme DdeI or isoschizomers of the restriction enzyme DdeI, wherein (a) is the primer pair composed of two lines of single-stranded DNA molecules shown in a sequence 1 and a sequence 2 in a sequence table, (b) is the primer pair which is composed of two lines of single-stranded DNA molecules shown in the sequence obtained by replacing and / or deleting and / or adding one or several nucleotides shown in the sequence 1 and the sequence 2 in the sequence table and is identical with the primer pair in (a) in function. According to the method, cytochrome b gene sequences of the traditional Chinese medical material corn cervi pantotrichum and counterfeit drug of the traditional Chinese medical material corn cervi pantotrichum are analyzed, different SNP loci of the traditional Chinese medical material corn cervi pantotrichum and the counterfeit drug of the traditional Chinese medical material corn cervi pantotrichum are obtained, identification primers and the restriction enzyme are designed, and the traditional Chinese medical material corn cervi pantotrichum and the counterfeit drug of the traditional Chinese medical material corn cervi pantotrichum are accurately identified through the restriction fragment length polymorohism polymerase chain reaction technology.

Description

Technical field [0001] The invention belongs to the field of identification of Chinese medicinal materials, and relates to a method for identifying Chinese medicinal antler based on PCR-RFLP, in particular to a kit for identifying Chinese medicinal antler based on PCR-RFLP and an identification method thereof. Background technique [0002] There are two sources for the basic source of velvet antler, deer CervusnipponTemminck or CervuselaphusLinnaeus, the unossified and densely hairy young horns of stags. The former is often called "flower antler" and the latter is called "red antler". Because deer antler is a fine and expensive Chinese medicinal material and its price is high, fake deer antler can often be seen in the medicinal material market at present, and the quality problem is serious. The main fake antlers include unossified antlers such as reindeer, moose, sambar, elk, roe deer, and small muntjac. Among them, reindeer, sambar, roe deer, etc. are more common. At present, ...

AI Technical Summary

Problems solved by technology

At present, people mainly distinguish velvet antler and its counterfeit products through traditional trait identification methods, but after processing, the traits and characteristics of velvet antler and its counterfeit products disappear, and the difference is not obvious

In addition, some studies have shown that the two strains can be identified using DNA sequencing technology and site-specific PCR technology, but the identification results of DNA sequencing technology are not intuitive enough, and the operation is slightly complicated

The use of site-specific PCR technology generally requires the design of a pair of primers for sika deer and red deer, which are generally only used to identify antler and other species such as cattle, yaks, sheep, etc. Differentiate between sika deer and red deer

On the other hand, DNA sequencing technology and site-specific PCR technology can only detect whether the sample to be tested contains genuine antler (sika deer or red deer), and cannot judge whether it is genuine antler or a mixed sample of genuine antler and counterfeit

[0003] The velvet velvet that circulates in the market has been processed, transported and stored, and even sliced ​​and sorted, its shape has changed and it is difficult to distinguish

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