Application of DNA Molecular Identification Method to Distinguish Ejiao and its Adulterants

Jing Wang (Wuhan Institute for Drug and Medical Device Control)

Abstract


To identify Ejiao and its adulterants at the DNA level by using DNA molecula marker. Ejiao (Asini Corii Colla) is a commonly used medicinal material. However, its adulteration is a serious concern. Due to the morphological characteristics of Asini Corii Colla and its adulterants, traditional identification techniques often complex and professional, which is not conducive to the circulation management and safety of the medicinal materials. To improve the distinction between Asini Corii Colla and its adulterants accurately, this study identified and its adulterant samples based on the CytB sequence. Sequence characteristics, Basic Local Alignment Search Tool (BLAST) application, genetic distance, construction of phylogenetic tree showed the CytB sequence to accurately identify Asini Corii Colla from its adulterants. Furthermore, in this study, we designed a specific primer, based on the CytB sequence, and established a PCR detection system for rapid, sensitive, and specific identification of Asini Corii Colla. Compared to DNA barcoding technology, this method has shorter detection time, stronger specificity, and higher sensitivity, which lays the foundation for the rapid identification of Asini Corii Colla.


Keywords


Ejiao; Asini Corii Colla; CytB; DNA barcoding technology

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References


[1] Wen-long Li, Hai-fan HAN, et al. Manufacturer identication and storage time determination of “Dong’e Ejiao” using near infrared spectroscopy and chemometrics. Journal of Zhejiang University-SCIENCE B (Biomedicine & Biotechnology), 2016 17(5): 382-390.

[2] Xiao L, Liao F, Zhou X, Qin Y, Miwa N. Biological effects of traditional Chinese medicine Colla corii asini. New Food Indust, 2019 61:6972.

[3] Jiao ZS, Wang JE, Li JT. Study on extraction processes for preparation of Colla Corii Asini. Chinese Traditional Patent Medicine, May, 2000, Vol. 22, No. 5.

[4] Chinese Pharmacopoeia. 2015 edition. (Ministry of Public Health of the People’s Republic of China, 2015).

[5] Li DZ, Liu JQ, et al. Plant DNA barcoding in China. Journal of Systematics and Evolution, 2011, 49 (3), 165-168.

[6] Liu YM. Study on methods of origin identification of traditional Chinese Medicine. Journal of Hubei University of Chinese Medicine, Dec 2014, Vol. 16, No. 6.

[7] Chen SL. Standard DNA barcodes of Chinese Materia Medica in Chinese Pharmacopoeia. Science Press. 2015.

[8] Kress WJ, Erickson DL. DNA barcodes: genes, genomics, and bioinformatics. Proc Natl Acad Sci USA, 2008,105:2761-2762.

[9] Chen SL, Yao H, Han JP, et al. Validation of the ITS2 region as a novel DNA barcode for identifying medicinal plant species. PloS ONE, 2010, 5: E8613.

[10] Hebert PDN, Cywinska A, Ball SL, et al. Biological identifications through DNA barcodes. Proc R Soc Biol Sci Ser B, 2003, 270: 313-321.

[11] Irwin D M, Kocher T D, Wilson A C. Evolution of cytochrome b gene of mammals. J Mol Evol, 1999, 32(2): 128-144.

[12] Kocher T D, Thomas W K, Meyer A, Edwards S V, Pặặbo S, Villablanca F X, Wilson A C. Dynamics of mitochondrial DNA evolution in animals: Amplification and sequencing with conserved primers. Proc Natl Acad Sci USA, 1989, 86(16): 6196-6200.

[13] Vladimir O A, Poltoraus L A, Zhivotovsky V. Mitochondrial DNA sequence diversity in Russians. In: FEBS Lett, 1999, 197-201.

[14] Wang YC, Li Na, et al. Comparison study on DNA extraction methods of deep procession products of Asini Corii Colla. Food and Drug, 2018, Vol. 20, No. 3.



DOI: https://doi.org/10.30564/jams.v4i2.2952

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