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A kind of specific primer, kit and identification method for identifying traditional Chinese medicine gecko

A kit and a specific technology, applied in the field of identification of traditional Chinese medicinal materials, can solve the problems of disappearance of characteristics and characteristics of fake products, unqualified products, difficult identification, etc., and achieve the effects of small molecular weight, high sensitivity and accurate identification results.

Active Publication Date: 2019-07-26
BEIJING JIANSHENG PHARMA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For some of these varieties, the morphological characteristics of Shougong authentic products are slightly different from those of Shougong. It is difficult to identify Shougong medicinal materials with traditional identification methods. In view of the changes in the morphology of Shougong in various links such as processing, processing, storage and transportation, The traits and characteristics of the gecko and its counterfeit products disappear, thus increasing the difficulty of identification
Driven by economic interests, the market phenomenon of "substituting fakes for real", "substituting substandard products" and "substandard products" are ubiquitous. accurately identify
In order to ensure the quality and clinical efficacy of Chinese herbal medicines, some studies have used modern molecular biology PCR amplification technology, which can complete the identification of some Chinese herbal medicines in a short time. Specific primers and methods for rapid identification of gecko species in traditional Chinese medicine

Method used

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  • A kind of specific primer, kit and identification method for identifying traditional Chinese medicine gecko
  • A kind of specific primer, kit and identification method for identifying traditional Chinese medicine gecko

Examples

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Effect test

Embodiment 1

[0042] This example provides a specific primer for identifying the gecko of traditional Chinese medicine. The specific primers designed for the CYT B gene of the gecko are as follows, as shown in SEQ ID NO: 1-2 in the sequence listing:

[0043] Upstream primer: 5′-CAACGGGGCCCTATATTCCG-3′;

[0044] Downstream primer: 5'-AGGGTTGCGTATGGGGTTTC-3'.

[0045] The above primers were synthesized by Shanghai Yingwei Jieji Company (Beijing Synthetic Department).

Embodiment 2

[0047] The present embodiment provides a kind of kit that is used for the identification of traditional Chinese medicine gecko, and each said kit includes the primer solution part and the PCR reaction solution part of independent packaging respectively; The specific primer; the PCR reaction solution part contains Mg-containing for PCR amplification 2+ Amplification buffer, dNTPs, Taq DNA polymerase, and ddH 2 O, the formula of each described test kit is:

[0048] Upstream primer, 10 μM, 1 μL;

[0049] Downstream primer, 10 μM, 1 μL;

[0050] PCR SuperMix, 10 μL;

[0051] wxya 2 O, 8 μL.

Embodiment 3

[0053] This embodiment provides a method for identifying the traditional Chinese medicine gecko using the specific primer of embodiment 1 or the kit of embodiment 2, comprising the following steps:

[0054] (1) Medicinal materials were collected in Shanxi, Anhui, Guangxi, Hebei and other provinces (Table 1). All the samples tested in Table 1 are commercially available products, which can be obtained through purchase, or collected or purchased from the source.

[0055] Table 1 Shougong and its common counterfeit sample sources

[0056]

[0057] Select the lyophilized powder of the sample in the above table 1 to adopt Genomic DNAKit extracts total DNA, and operates according to the instructions, the steps are as follows:

[0058] (1) extract the total DNA of the Chinese medicine gecko to be tested

[0059] 1) Take the freeze-dried powder of each sample ≤ 25mg, and place it in a sterile 1.5mL centrifuge tube;

[0060] 2) Add 100 μL of LB2 (lysate) and 20 μL of Proteinase K...

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Abstract

The invention provides a specific primer, a kit and an identification method for identifying traditional Chinese medicine kwangsi gecko. A lot of primers are designed based on a CYTB gene of the kwangsi gecko; the primer with species specificity is screened out through the screening study of a large amount of primers; the primer is used for identifying the traditional Chinese medicine kwangsi gecko, so that the kwangsi gecko medicinal material can be accurately separated from other adulterants, a PCR (Polymerase Chain Reaction) amplification product is only 100 to 250bp, the molecular weight is low, a lot of traditional Chinese medicine kwangsi gecko is ensured to be identified within a short time, and an identification result is accurate, stable, reliable and high in sensitivity.

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 gecko of traditional Chinese medicine. Background technique [0002] Gekko Swinhoana Gunther is the whole Gekko Swinhoana Gunther. Its medicine is cold in nature, salty in taste, slightly poisonous, and enters the lung and kidney meridians. It has the effects of dispelling stagnation and detoxification, nourishing yin and cooling blood, quenching wind and spasm, and penetrating meridians and collaterals. There are many records about ghougong in ancient medical classics. For example, in "Compendium of Materia Medica", ghougong is mainly used for stroke paralysis, immobility of hands and feet, or chronic wind pain, epilepsy due to wind convulsion, infantile malnutrition, blood accumulation scrofula, treatment Scorpion sting; Zhao Qiguang in the Qing D...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6888
Inventor 李建生
Owner BEIJING JIANSHENG PHARMA
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