Multiplex Allele-Specific PCR Method for Identification of Syngnathus and Its Adulterants

LIU Fu-yan; JIN Yan; YUAN Yuan; QIN Wen; ZHAO Yu-yang; JIANG Chao

doi:10.13422/j.cnki.syfjx.20181136

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Multiplex Allele-Specific PCR Method for Identification of Syngnathus and Its Adulterants

更新时间：2024-01-04

- Multiplex Allele-Specific PCR Method for Identification of Syngnathus and Its Adulterants
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 24, Issue 15, Pages: 57-64(2018)
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- DOI：10.13422/j.cnki.syfjx.20181136
  CLC： R282.5
- Published：2018
- 稿件说明：
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LIU Fu-yan, JIN Yan, YUAN Yuan, et al. Multiplex Allele-Specific PCR Method for Identification of Syngnathus and Its Adulterants[J]. Chinese journal of experimental traditional medical formulae, 2018, 24(15): 57-64.
DOI：

LIU Fu-yan, JIN Yan, YUAN Yuan, et al. Multiplex Allele-Specific PCR Method for Identification of Syngnathus and Its Adulterants[J]. Chinese journal of experimental traditional medical formulae, 2018, 24(15): 57-64. DOI： 10.13422/j.cnki.syfjx.20181136.

摘要

目的：建立一种高效、准确的中药材海龙及其常见混伪品的特异性聚合酶链式反应（polymerase chain reaction，PCR）鉴别方法。方法：通过比较海龙及其混伪品的细胞色素C氧化酶亚基Ⅰ基因（cytochrome C oxidase subunit Ⅰ，COⅠ）基因序列差异，根据变异位点设计刁海龙、尖海龙及拟海龙的特异性鉴别引物，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。在此基础上，将3对特异性引物组合，构建多重PCR体系。结果：在多重PCR体系中，刁海龙能扩增出485 bp片段，尖海龙可扩增出120 bp片段，拟海龙可以扩增出240 bp片段，其他混伪品均无条带。所设计的特异性鉴别引物具有高度的特异性。结论：该文所建立的位点特异性PCR鉴别方法可实现海龙的准确鉴别。

Abstract

Objective: To establish an effective and accurate specific polymerase chain reaction (PCR) identification method for herb Syngnathus and its common adulterates. Method: Based on the difference in cytochrome C oxidase subunit Ⅰ(COⅠ) gene DNA sequences among Solenognathus hardwickii

Syngnathoides biaculeatus

S. acus and adulterants

the specific primers were designed; the reaction conditions were optimized

and the PCR method for identification was explored and verified in terms of tolerance and feasibility. The three pairs of specific primers were combined to build multiplex PCR systems. Result: Through the established multiplex PCR reaction system

485

240

120 bp fragments were amplified from DNA templates of S. shardwickii

S. biaculeatus and S. acus

respectively. All the adulterants had no bands. Conclusion: The designed identification primers were highly specific

and the PCR amplification of specific alleles established in this paper can be used to accurately identify the Syngnathus.

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Related Author

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Yu-yang ZHAO

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Related Institution

College of Traditional Chinese Medicine, Guangdong Pharmaceutical University

State Key Laboratory Breeding Base of Dao-di Herbs, China Academy of Chinese Medical Sciences, National Resource Center for Chinese Materia Medica

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School of Pharmacy，Anhui University of Chinese Medicine

School of Traditional Chinese Pharmacy，China Pharmaceutical University

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