Specific PCR Method for Identification of Medicinal Scolopendra

Na TIAN; Yuan YUAN; Yan JIN; Quan YANG; Yu-yang ZHAO; Chao JIANG

doi:10.13422/j.cnki.syfjx.20191711

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Specific PCR Method for Identification of Medicinal Scolopendra

Topic on DNA Molecular Identification of Animal Medicinal Materials | 更新时间：2021-02-09

- Specific PCR Method for Identification of Medicinal Scolopendra
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 25, Issue 17, Pages: 113-117(2019)
- 作者机构：
  
  1.广东药科大学　中药学院，广州　 510006；
  2.中国中医科学院　中药资源中心，道地药材国家重点实验室培育基地，北京　 100700
- 作者简介：
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.20191711
  CLC： R284.2; R285; R22; R2-031
- Received：04 January 2019，
  
  Published Online：17 May 2019，
  
  Published：05 September 2019
- 稿件说明：
移动端阅览
Na TIAN, Yuan YUAN, Yan JIN, et al. Specific PCR Method for Identification of Medicinal Scolopendra[J]. Chinese journal of experimental traditional medical formulae, 2019, 25(17): 113-117.
DOI：

Na TIAN, Yuan YUAN, Yan JIN, et al. Specific PCR Method for Identification of Medicinal Scolopendra[J]. Chinese journal of experimental traditional medical formulae, 2019, 25(17): 113-117. DOI： 10.13422/j.cnki.syfjx.20191711.

摘要

目的：

蜈蚣是我国传统的中药材，具有良好的药用价值。目前市场上蜈蚣的混伪品日益增多，临床用药的安全性和有效性难以保证。该研究建立了一种蜈蚣基原物种的特异性聚合酶链式反应(PCR)方法，以准确、简便的鉴别蜈蚣及其常见混伪品。

方法：

通过比较蜈蚣及其混伪品基原物种的COI基因序列差异，根据变异位点设计蜈蚣正品少棘巨蜈蚣的特异性鉴别引物WG-500.F和WG-500.R，采集蜈蚣原动物样本及药材样本，优化反应体系并对此方法进行耐受性和适用性的考察和验证。

结果：

少棘巨蜈蚣原动物样本及蜈蚣正品药材使用设计的蜈蚣鉴别引物经PCR扩增和凝胶电泳后出约500 bp的单一明亮条带，其他混伪品如多棘蜈蚣、模棘蜈蚣、哈氏蜈蚣、海南蜈蚣等均无条带出现。

结论：

PCR鉴别方法可准确鉴别蜈蚣原动物和药材的真伪，具有高度特异性，为中药材蜈蚣的真伪鉴别提供了良好的科学依据。该方法具有简单、直观等特点，有利于广泛的普及与应用，在中药材鉴定方面有着广阔的应用前景。

Abstract

Objective:

Scolopendra was a traditional Chinese medicine(TCM) with a good medicinal value. Nowadays

there have been increasingly more adulterates of Scolopendra in the medicine market. To ensure the safe and effectiveness of clinical medicines

a convenient and accurate specific polymerase chain reaction(PCR) method for identification of medicinal Scolopendra from its common adulterates was established.

Method:

Based on the differences of COI gene DNA sequences among

Scolopendra subspinipes mutilans

and adulterants

the specific primer was designed

the reaction conditions were optimized

and the PCR method for identification was explored and verified in terms of tolerance and feasibility. Besides

the original animal samples and medicine of Scolopendra were collected.

Result:

Through the established PCR reaction system

the bright and simple fragments of 500 bp was amplified from DNA templates of

subspinipes mutilans

. All of the adulterants were negative by the multiplex PCR assay

such as

multidens

subspinipes

dehaani

hainanum

Conclusion:

The identified primer is highly specific

and the specific PCR method established in this paper can accurately identify Scolopendra and its adulterants

so as to provide an excellent scientific basis for the identification of TCM Scolopendra. The method is simple and intuitive

and facilitates wide promotion and application of the method

with a broad application prospect in the identification of TCM.

关键词

Keywords

references

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