Shokralla Shadi, Hellberg Rosalee S, Handy Sara M, King Ian, Hajibabaei Mehrdad
Biodiversity Institute of Ontario and Department of Integrative Biology, University of Guelph, Guelph, ON N1G 2W1, Canada.
Department of Microbiology, Mansoura University, Mansoura 35516, Egypt.
Sci Rep. 2015 Oct 30;5:15894. doi: 10.1038/srep15894.
Species substitution is a form of seafood fraud for the purpose of economic gain. DNA barcoding utilizes species-specific DNA sequence information for specimen identification. Previous work has established the usability of short DNA sequences-mini-barcodes-for identification of specimens harboring degraded DNA. This study aims at establishing a DNA mini-barcoding system for all fish species commonly used in processed fish products in North America. Six mini-barcode primer pairs targeting short (127-314 bp) fragments of the cytochrome c oxidase I (CO1) DNA barcode region were developed by examining over 8,000 DNA barcodes from species in the U.S. Food and Drug Administration (FDA) Seafood List. The mini-barcode primer pairs were then tested against 44 processed fish products representing a range of species and product types. Of the 44 products, 41 (93.2%) could be identified at the species or genus level. The greatest mini-barcoding success rate found with an individual primer pair was 88.6% compared to 20.5% success rate achieved by the full-length DNA barcode primers. Overall, this study presents a mini-barcoding system that can be used to identify a wide range of fish species in commercial products and may be utilized in high throughput DNA sequencing for authentication of heavily processed fish products.
物种替代是一种以获取经济利益为目的的海鲜欺诈形式。DNA条形码技术利用物种特异性的DNA序列信息来鉴定标本。先前的研究已经证实了短DNA序列——微型条形码——在鉴定含有降解DNA的标本方面的可用性。本研究旨在为北美加工鱼产品中常用的所有鱼类建立一个DNA微型条形码系统。通过检查美国食品药品监督管理局（FDA）海鲜清单中物种的8000多个DNA条形码，开发了针对细胞色素c氧化酶I（CO1）DNA条形码区域短片段（127 - 314bp）的六对微型条形码引物对。然后，将这些微型条形码引物对应用于44种代表一系列物种和产品类型的加工鱼产品进行测试。在这44种产品中，41种（93.2%）能够在物种或属的水平上得到鉴定。与全长DNA条形码引物20.5%的成功率相比，单个引物对的最大微型条形码成功率为88.6%。总体而言，本研究提出了一种微型条形码系统，可用于鉴定商业产品中的多种鱼类物种，并可用于高通量DNA测序以鉴定深加工鱼产品的真伪。
