Quality Evaluation of Galli Gigerii Endothelium Corneum by Allele-specific PCR Method

Jing-zhe PU; Ya-zhong ZHANG; Ye-lin ZHU; Chao JIANG; Yuan YUAN

doi:10.13422/j.cnki.syfjx.20191717

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Quality Evaluation of Galli Gigerii Endothelium Corneum by Allele-specific PCR Method

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

- Quality Evaluation of Galli Gigerii Endothelium Corneum by Allele-specific PCR Method
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 25, Issue 17, Pages: 142-147(2019)
- 作者机构：
  
  1.安徽省食品药品检验研究院，合肥　 230051；
  2.安徽省市场监督管理局，合肥　 230051；
  3.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.20191717
  CLC： R284.2; R285; R22; R2-031
- Received：19 March 2019，
  
  Published Online：11 June 2019，
  
  Published：05 September 2019
- 稿件说明：
移动端阅览
Jing-zhe PU, Ya-zhong ZHANG, Ye-lin ZHU, et al. Quality Evaluation of Galli Gigerii Endothelium Corneum by Allele-specific PCR Method[J]. Chinese journal of experimental traditional medical formulae, 2019, 25(17): 142-147.
DOI：

Jing-zhe PU, Ya-zhong ZHANG, Ye-lin ZHU, et al. Quality Evaluation of Galli Gigerii Endothelium Corneum by Allele-specific PCR Method[J]. Chinese journal of experimental traditional medical formulae, 2019, 25(17): 142-147. DOI： 10.13422/j.cnki.syfjx.20191717.

摘要

目的：

建立鉴别鸡内金及其常见伪品的物种特异性聚合酶链式反应(PCR)方法，对全国中药材及饮片专项抽验任务中所抽取的鸡内金饮片进行真伪鉴别，评价市场上鸡内金的真伪情况。

方法：

根据鸡、鸭、鹅的12S序列差异，设计或优化物种特异性PCR鉴别引物，对影响PCR结果的主要因素退火温度，PCR循环次数，模板DNA浓度，

Taq

酶种类等进行方法学考察和优化。使用优化的特异性PCR方法对鸡、鸭、鹅内金样品进行DNA分子鉴别。

结果：

在进行方法学考察确定最优鉴别条件的基础上，当PCR退火温度为55 ℃，循环次数为30次的条件下，当使用文中筛选得到的鸡物种特异性鉴别引物时，所测鸡内金饮片均检出约为273 bp的特异性扩增条带，混伪品鸭内金和鹅内金均无相应扩增条带；当使用鸭和鹅鉴别引物时，均未检出相应的扩增条带。

结论：

位点特异性PCR方法能够快速准确的鉴别出鸡内金真伪品，可用于全国中药材及饮片专项抽验任务中鸡内金的真伪鉴定，目前市场上用鸭内金和鹅内金充鸡内金现象较少，质量评价较好。

Abstract

Objective:

To screen the specific reverse primers of Galli Gigerii Endothelium Corneum

duck gizzard membrane and goose gizzard membrane

and establish a specific PCR for molecular identifying Galli Gigerii Endothelium Corneum and its common adulterants.

Method:

Based on the mutation sites on the 12S rRNA sequence

specific polymerase Chain reaction(PCR) identify primers were designed for chicken

duck and goose gizzard membrane. The specific PCR reaction conditions were optimized

and the PCR identification method was explored and verified in terms of tolerance and feasibility. Thirty batches of Galli Gigerii Endothelium Corneum decoction pieces extracted from the test were identified.

Result:

Thirty batches of Galli Gigerii Endothelium Corneum decoction pieces were detected using chicken-specific primers

273 bp of specific bands was amplified and visualized on the agarose electrophoregram. When duck and goose primers were used

no corresponding amplified band was detected.

Conclusion:

The allele-specific PCR method can be used as a rapid and accurate method to identify Galli Gigerii Endothelium Corneum. It is a promise method for special sampling tasks of Chinese herbal medicine and decoction tablets nationwide.

关键词

Keywords

references

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