Rapid detection of CITES-listed shark fin species by loop-mediated isothermal amplification assay with potential for field use

Grace Wing-Chiu But,Kwang-Tsao Shao,Pang-Chui Shaw,Hoi-Yan Wu

doi:10.1038/s41598-020-61150-8

Abstract

Shark fin is a delicacy in many Asian countries. Overexploitation of sharks for shark fin trading has led to a drastic reduction in shark population. To monitor international trade of shark fin products and protect the endangered species from further population decline, we present rapid, user-friendly and sensitive diagnostic loop-mediated isothermal amplification (LAMP) and effective polymerase chain reaction (PCR) assays for all twelve CITES-listed shark species. Species-specific LAMP and PCR primers were designed based on cytochrome oxidase I (COI) and NADH2 regions. Our LAMP and PCR assays have been tested on 291 samples from 93 shark and related species. Target shark species could be differentiated from non-target species within three hours from DNA extraction to LAMP assay. The LAMP assay reported here is a simple and robust solution for on-site detection of CITES-listed shark species with shark fin products.

Highlights

Shark fin is a delicacy in many Asian countries
We have developed sensitive, simple to use, and to our knowledge, the fastest species-specific assays using loop-mediated isothermal amplification (LAMP) technique, as well as simple and low-cost polymerase chain reaction (PCR) assays to authenticate all 12 CITES-listed shark species
The LAMP and PCR assays we developed can successfully discriminate their target species from the other 92 shark and related species, which has included species commonly found in the Hong Kong shark fin market[12]

Summary

Introduction

Shark fin is a delicacy in many Asian countries. Overexploitation of sharks for shark fin trading has led to a drastic reduction in shark population. To monitor international trade of shark fin products and protect the endangered species from further population decline, we present rapid, user-friendly and sensitive diagnostic loop-mediated isothermal amplification (LAMP) and effective polymerase chain reaction (PCR) assays for all twelve CITES-listed shark species. Regular practices of molecular means for shark species identification include DNA barcoding[3,14,15], mini-DNA barcoding for highly degraded DNA16, and species-specific amplification with PCR-based method[17,18,19,20,21,22] These genetic approaches for identifying the species-of-origin of shark fins have been carried out www.nature.com/scientificreports since 1990s and they are effective for shark fin samples with different levels of processing. Recent studies have applied the LAMP technique on species-specific detection of animals[27,28] and medicinal material[29,30]

Methods

Results

Conclusion