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Primer group and crude drug identification method using primer group

A primer set and crude drug technology, applied in biochemical equipment and methods, microbial measurement/inspection, recombinant DNA technology, etc., can solve the problems of inability to accurately identify the original plant and difficulty in determining the base sequence

Pending Publication Date: 2020-07-07
TSUMURA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, it is very difficult to determine the base sequence of the nucleic acid collected from the crude drug itself, and it has not been possible to accurately identify the original plant so far.

Method used

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  • Primer group and crude drug identification method using primer group
  • Primer group and crude drug identification method using primer group
  • Primer group and crude drug identification method using primer group

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0241]

[0242] 1 Introduction

[0243] In the seventeenth revised edition of the Japanese Pharmacopoeia, the original plant of Poria cocos is Wolfiporia cocos Ryvarden et Gilbertson (Poria cocos Wolf) (Poria cocos Wolf) (Poria cocos Wolf). However, it is well known that there are many kinds of internal mutations in the same species. Although there are varieties in the Japanese market that meet the regulations of the Japanese Pharmacopoeia, it is also possible to circulate multiple lines with different base sequences. The ingredients contained in different strains may be different. In order to ensure the quality of the medicine, it is an effective way to avoid mixing these different strains.

[0244] Therefore, the present inventors conducted a comprehensive study on the genomic DNA of various Poria strains and their related bacteria. The results showed that in the sequence of the ITS region in the ribosomal DNA, Poria, a major circulating product in the market, has a fixed sequen...

Embodiment 2

[0285]

[0286] 1 Introduction

[0287] In the seventeenth revised edition of the Japanese Pharmacopoeia, the original plant of Rehmannia glutinosa is Rehmanniaglutinosa Liboschitz var.purpurea Makino or Rehmannia glutinosa Liboschitz (Scrophulariaceae). However, other closely related species such as Rehmannia henryi may be circulated in the market. In order to ensure the quality of drugs, the key is to avoid misuse and mixing them.

[0288] Therefore, the inventors conducted a comprehensive study on the genomic DNA or chloroplast DNA of Rehmannia glutinosa and its relatives. The results showed that in the sequence of ITS region in ribosomal DNA, Rehmannia glutinosa has a fixed sequence in this region, which can be distinguished from other related species. Therefore, this region is a useful DNA marker for distinguishing Rehmannia glutinosa from other related species.

[0289] However, it is often difficult to determine the sequence from crude drugs due to the following reasons: 1) ...

Embodiment 3

[0314]

[0315] 1 Introduction

[0316] In the seventeenth revised edition of the Japanese Pharmacopoeia, the original plant of Corydalis is Corydalisturtschaninovii Besser forma yanhusuo Y.H.Chou et C.C.Hsu (Papaveraceae). However, other related species, Corydalis ambigua, may be circulated in the market. In order to ensure the quality of the drug, the key is to avoid misuse and mixing them.

[0317] Therefore, the inventors conducted a comprehensive study on the genomic DNA or chloroplast DNA of Corydalis and its relatives. The results showed that in the sequence of the ITS region of ribosomal DNA, Corydalis has a fixed sequence in this region, which can be distinguished from other related species. Therefore, this region is a useful DNA marker for distinguishing Yanhusuo from other related species.

[0318] However, it is often difficult to determine the sequence from crude drugs due to the following reasons: 1) DNA damage and fragmentation caused by time changes and processing; ...

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Abstract

An aim of the present invention is to develop and provide a means for accurately identifying whether or not a plant to be detected, which is a candidate crude drug after processing, is a base plant, and identifying the candidate crude drug as a target crude drug and an identification method using the means. For various crude drugs, a primer group capable of stably acquiring base sequence information from a processed to-be-detected plant and amplifying a specific region in genome DNA or chloroplast DNA of a base plant with a specific base sequence in the base plant is designed. The invention provides the primer group and a crude drug identification method using the primer group.

Description

Technical field [0001] The present invention relates to a nucleic acid amplification reaction primer set capable of confirming that various crude drug samples are the Japanese Pharmacopoeia or the crude drug standards other than the Japanese Pharmacopoeia (hereinafter referred to as "exigen regulations"). Identification method of crude drugs using this primer set. Background technique [0002] Regarding crude drugs, in the seventeenth revised edition of the Japanese Pharmacopoeia or Outer Regulations 2015, the original plant is specified. Primitive plants refer to plants (protoplants) used as raw materials for a certain crude drug, and each crude drug is specified on a species basis. For example, the base plant of the crude drug "Rehmannia glutinosa" is Rehmannia glutinosa Liboschitzvar.purpurea Makino or Rehmannia glutinosa Liboschitz (Scrophulariaceae). Plant species other than this are not recognized in principle, even if they are related species of the same genus. For "rehm...

Claims

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Application Information

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IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895
Inventor 司马真央山路弘树曾根美佳子
Owner TSUMURA
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