Specific DNA mini-barcoding for identification of Gekko gecko and its products
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Chinese Medicine Open Access
RESEARCH
Specific DNA mini‑barcoding for identification of Gekko gecko and its products Yanyan Su1,2†, Dandan Ding1†, Mengjie Yao3, Lan Wu1, Gangqiang Dong2, Dong Zhang1, Shilin Chen1* and Li Xiang1*
Abstract Background: The dry body of the Tokay Gecko (Gekko gecko) is the source of a valuable traditional Chinese medicine, it is therefore listed as a Class II protected animal species in China. Due to increasing market demand and a declining supply of the species, a considerable number of adulterants have emerged in the market. Thus, it is necessary to establish an accurate and rapid method of identification for distinguishing G. gecko from its adulterants and for separating it from highly processed products. Methods: A total of 274 COI sequences were analyzed by using MEGA 5.0 software. Several specific primers were designed to amplify mini-barcode regions and identify G. gecko from its counterfeits and products. Results: 274 COI sequences of G. gecko and 15 adulterants species were analyzed. G. gecko could be distinguished from its adulterants through BLAST analysis, intra- and inter-specific distance analyses, and an NJ tree based on COI sequences. Two pairs of specific primers designed for this study, COISF2/COISR2 and COISF3/COISR3, amplified 200and 133-bp fragments of the COI region, respectively, both of which were suitable for the identification of G. gecko and its adulterants. Furthermore, COISF3/COISR3 detected G. gecko in 15 batches of products. Conclusion: Therefore, the specific DNA mini-barcoding method developed here may be a powerful tool for the identification of G. gecko and counterfeits, and may also be used to distinguish G. gecko from its highly processed by-products. Keywords: Gekko gecko, specific primers, DNA mini-barcoding, Identification Background As one of the rarest Chinese medicinal materials, the dry body of Tokay Gecko (G. gecko Linnaeus), has been used for two thousand years and is known for its remarkably therapeutic effect on kidney deficiency [1, 2]. Previous studies have found that G. gecko also has notable effects in relieving asthma, strengthening the immune system, *Correspondence: [email protected]; [email protected] † Yanyan Su and Dandan Ding contributed equally to this work 1 Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China Full list of author information is available at the end of the article
and treating tumors [3–5]. Today it is widely used in many functional foods and Chinese patent medicines, such as tinctures, Renshen Gejie Powder, Gejie Dingchuan Capsule, Gejie Dingchuan Pill, and Shenge Pingchuan Capsule [6, 7]. For decades, the medicinal demand for G. gecko has skyrocketed in China. However, due to the destruction of its natural habitats and indiscriminate hunting, the wild G. gecko has been classified as a Class II protected animal species in China since 1989 [8]. As a result, the demand for the animal is considerably higher than its supply. Currently, the domestic market rel
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