Chinese Journal of Pharmaceutical Analysis >

2023 , Vol. 43 >Issue 7: 1263 - 1269

DOI: https://doi.org/10.16155/j.0254-1793.2023.07.21

Rapid Analysis

Authentication of Agkistrodon acutus based on loop-mediate isothermal amplification*

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  • 1. College of Nursing and Health Management, Lingnan Institute of Technology, Guangzhou 510663, China;
    2. Institute of Analysis, Guangdong Academy of Sciences (China National Analytical Center Guangzhou), Guangzhou 510070, China;
    3. School of Pharmacy, Lingnan Institute of Technology, Guangzhou 510663, China;
    4. Pharmacy Department of Maoming people's Hospital, Maoming 525000, China

Received date: 2023-06-16

  Online published: 2024-06-24

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Abstract

Objective: To establish a rapid visual identification method for Agkistrodon acutus based on loop-mediated isothermal amplification (LAMP). Methods: The LAMP assay for the detection of Agkistrodon acutus was developed by designing four primers including two inner primers and two outer primers which according to the specific sequence of Agkistrodon acutus contained in the Chinese Pharmacopoeia. Specificity and sensitivity of the LAMP assay was validated by comparing the polymerase chain reaction (PCR) method included in the Chinese Pharmacopoeia, and using the LAMP assay and the PCR to identify the authenticity of agkistrodon acutus on the market. Results: The LAMP assay established by using those primers showed good performance on specificity and sensitivity. Amplification could be observed in reference crude drug of Agkistrodon acutus and four samples of Agkistrodon acutus from the market,and there was no amplification in one sample of Agkistrodon acutus from the market and eight common adulterants, which was consistent with the identification results of PCR. The sensitivity of the LAMP assay was 0.004 ng·μL-1 and the sensitivity of the PCR assay was 0.4 ng·μL-1, so that the LAMP assay showed higher sensitivity. Conclusion: The result demonstrated that the specific LAMP assay can be used for rapid detection of Agkistrodon acutus.

Key words： Agkistrodon acutus; LAMP; nucleic acid isothermal amplification technology; polymerase chain reaction; identification of traditional Chinese medicine; molecular detection; rapid identification

Cite this article

DENG Hua-ming, XU Xiao-ke, HUANG Qi-hong, LI Zi-hang, LIANG Lin-yuan, YU Zhi-xing, LIANG Yong-heng . Authentication of Agkistrodon acutus based on loop-mediate isothermal amplification*[J]. Chinese Journal of Pharmaceutical Analysis, 2023 , 43(7) : 1263 -1269 . DOI: 10.16155/j.0254-1793.2023.07.21

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