Araliaceae traditional Chinese medicine product identification kit and araliaceae traditional Chinese medicine product identification method

A technology of traditional Chinese medicine products and kits, which is applied in the direction of biochemical equipment and methods, DNA/RNA fragments, microbe determination/inspection, etc. It can solve the problems of low identification efficiency of ginseng, Panax notoginseng and American ginseng, and achieve good specificity Effect

Pending Publication Date: 2020-04-21
SPH XING LING SCI & TECH PHARM CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In view of the shortcomings of the prior art described above, the object of the present invention is to provide a kit and method for identifying Araliaceae traditional Chinese medicine products, which are used to solve the problem of low identification efficiency of ginseng, Panax notoginseng and American ginseng among traditional Chinese medicinal materials in the prior art

Method used

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  • Araliaceae traditional Chinese medicine product identification kit and araliaceae traditional Chinese medicine product identification method
  • Araliaceae traditional Chinese medicine product identification kit and araliaceae traditional Chinese medicine product identification method
  • Araliaceae traditional Chinese medicine product identification kit and araliaceae traditional Chinese medicine product identification method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] Example 1 Extraction of medicinal material genomic DNA

[0056] 1) Put the experimental materials after the surface sterilization treatment into a sterile mortar, add liquid nitrogen in an appropriate amount several times, and forcefully grind the plant tissue into powder;

[0057] 2) Transfer the powder to a sterile centrifuge tube, and add CTAB extraction buffer (2% CTAB; 100mM Tris-HCl pH8.0; 1.4M NaCl; 20mM EDTA; 1.5% polyvinyl-pyrrolidone, PVP; 0.5% 2-mercaptoethanol) inverted and mixed.

[0058] 3) Water bath at 60°C for 45 minutes, and mix by inverting several times during the period.

[0059] 4) Add an equal volume of chloroform:isoamyl alcohol (24:1), mix thoroughly, and centrifuge at 12000 rpm at 4°C for 10 min.

[0060] 5) Transfer the supernatant to a new sterile centrifuge tube, repeat step 4) once.

[0061] 6) Transfer the supernatant to a new sterile centrifuge tube, add an appropriate amount of RNase, and bathe in water at 37°C for 30 minutes.

[006...

Embodiment 2

[0066] Example 2 PCR amplification and agarose gel electrophoresis detection

[0067] (1) Primer design

[0068] Design genus-specific primers for ginseng, Panax notoginseng, and American ginseng, respectively:

[0069] a. Design primer pairs for the ginseng plant gene:

[0070] SEQ ID NO.1 forward primer: 5'-ATAACAATACCGGGCTGATAC-3';

[0071] SEQ ID NO.2 reverse primer: 5'-GCCAGTTAAGGACAGGAG-3';

[0072] b. Design primer pairs for the Panax notoginseng plant gene:

[0073] SEQ ID NO.3 forward primer: 5'-AGGGATGAGGGGTGCGTAG-3';

[0074] SEQ ID NO.4 reverse primer: 5'-CGACATGAGAAGAGGGCTTTTA-3';

[0075] c. Design primer pairs for American ginseng plant genes:

[0076] SEQ ID NO.5 forward primer: 5'-GATTTCTGCATATACGCCCAATT-3';

[0077] SEQ ID NO.6 reverse primer: 5'-GGAAAATGCGGGTTATGACAAA-3';

[0078] (2) PCR amplification

[0079] a. PCR reaction system

[0080] Using the genomic DNA contained in the sample to be tested obtained in Example 1 as a template, the three s...

Embodiment 3

[0089] Example 3 Ginseng, Panax notoginseng and American ginseng primers PCR amplification of single-species genomic DNA of ginseng, notoginseng and American ginseng

[0090] Genomic DNA of ginseng, Panax notoginseng, and American ginseng was diluted with ultrapure water to a concentration of 50 ng / μL, and used as a DNA template; a blank control was set, and 1 μL of ultrapure water was used to replace the DNA template.

[0091] (1) PCR amplification

[0092] The primers, PCR system and amplification program in Examples 1 and 2 were used for PCR amplification.

[0093] (2) Agarose gel electrophoresis detection

[0094] Take 10 μL of the PCR amplification product and add 1 μL of 10X loading buffer (containing 0.25% bromophenol blue, 0.25% xylene aniline, 60% glycerol), and use agarose gel electrophoresis with a concentration of 1.5% by mass percentage, voltage 180V, electrophoresis for 25min , photographed and observed under the gel imager, the results are shown in figure 1 ....

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PUM

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Abstract

The invention provides an araliaceae traditional Chinese medicine product identification kit and an araliaceae traditional Chinese medicine product identification method. The kit at least comprises the following three groups of primer pairs: a ginseng identification primer pair, wherein the forward primer is represented by SEQ ID NO:1, and the reverse primer is represented by SEQ ID NO:2; a pseudo-ginseng identification primer pair, wherein the forward primer is represented by SEQ ID NO:3, and the reverse primer is represented by SEQ ID NO:4; and an American ginseng identification primer pair,wherein forward primer is represented by SEQ ID NO:5, and the reverse primer is represented by SEQ ID NO:6. The kit and the method are mainly used for qualitatively identifying ginseng, pseudo-ginseng and American ginseng in traditional Chinese medicine products, and have high sensitivity and good specificity.

Description

technical field [0001] The invention relates to a kit and a method for identifying traditional Chinese medicinal products of Araliaceae. Background technique [0002] Traditional Chinese medicine has a special and important position in my country's medical and health undertakings, and Chinese herbal medicines are the foundation of the development of Chinese medicine. In recent years, the Chinese medicine industry has developed rapidly, service capabilities have been significantly enhanced, and the scale of the Chinese medicine industry has been significantly expanded. With the rapid development of the Chinese medicine industry and the growing demand for Chinese medicine both at home and abroad, Chinese medicinal materials have become an important resource for market competition. However, the development of traditional Chinese medicine is also facing many difficulties and problems. There are many recent products of some traditional Chinese medicinal materials, and the qualit...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895C12Q2600/16
Inventor 张亚光王丹丹陈丹青张艳欣
Owner SPH XING LING SCI & TECH PHARM CO LTD
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