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Rapid authentication of Eupolyphaga sinensis by loop-mediated isothermal amplification*

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  • 1. Guangdong Second Traditional Chinese Medicine Hospital (Guangdong Province Engineering Technology Research Institute of Traditional Chinese Medicine), Guangdong Provincial Key Laboratory of Research and Development in Traditional Chinese Medicine, Guangzhou, Guangdong 510095, China;
    2. The School of the Fifth Clinical Medicine, Guangzhou University of Chinese Medicine, Guangzhou 510405, China;
    3. School of Pharmacy, Guangzhou Xinhua University, Guangzhou 510520, China

Received date: 2024-01-18

  Online published: 2024-10-17

Abstract

Objective: To establish an authentication method for Eupolyphaga sinensis based on loop-mediated isothermal amplification (LAMP). Methods: Two pairs of primers (outer primer DB-F3,DB-B3 and inner primer DB-FIP,DB-BIP) were designed according to Eupolyphaga sinensis COⅠspecific loci. The reaction system containing template, primers, Bst DNA polymerase and methyl red-phenol red indicator amplified at 63 ℃ for 90 min. The LAMP results were determined by visual observation and compared with DNA barcoding and polymerase chain reaction (PCR) methods to investigate the specificity and sensitivity. Results: All 11 batches of the authentic Eupolyphaga sinensis sample tubes colour changed from purple red to orange yellow, while sample tubes of 1 batch of Steleophaga plancyi, 9 batches Opisthoplatia orientalis and 1 batch of the adulterant, Cybister tripunctatus orientalis, remained purple red after the reaction. The LAMP results were consistent with those of DNA barcoding and PCR. The detection limit of LAMP was 0.984 ng·mL-1. Conclusion: The established LAMP method is accurate, sensitive and easy to operate, low equipment required, and can be applied to the rapid screening of the authentication of Eupolyphaga sinensis.

Cite this article

CHEN Wei-tao, LI Yang-xue, JIANG Jie-yi, XIE Can-hui, JIA De-zheng, LIU Xiao-tong, LIAO Hui-na, LI Su-mei . Rapid authentication of Eupolyphaga sinensis by loop-mediated isothermal amplification*[J]. Chinese Journal of Pharmaceutical Analysis, 2024 , 44(9) : 1529 -1534 . DOI: 10.16155/j.0254-1793.2024-0035

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