Application of Loop-Mediated Isothermal Amplification (LAMP) for Rapid Detection of Jumbo Flying Squid Dosidicus gigas (
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Application of Loop-Mediated Isothermal Amplification (LAMP) for Rapid Detection of Jumbo Flying Squid Dosidicus gigas (D’Orbigny, 1835) Jian Ye 1 & Junli Feng 1 & Zhiyuan Dai 1 & Lu Meng 1 & Yanping Zhang 1 & Xiaona Jiang 1
Received: 5 May 2016 / Accepted: 17 October 2016 # Springer Science+Business Media New York 2016
Abstract Rapid identification of commercially important squid species is essential to manage squid market and protect consumers’ interests. In this study, two sets of loop-mediated isothermal amplification (LAMP)-based assays were developed for identification of jumbo flying squid (Dosidicus gigas). After analyzing the D. gigas mitochondrial genes reported in GenBank, the mitochondrial cytochrome c oxidase subunit I (COI) gene was selected for designing species-specific primers. When incubated at 65 °C for 30 min, only D. gigas DNA can be detected by real-time fluorescence detection (RealAmp) or visual detection via SYBR Green® I staining (Visual-LAMP). The detection limits were 10 pg per reaction for purified D. gigas DNA, and 0.01 % w/w D. gigas in homogenized cephalopod mixtures. These methods can be applied to all kinds of processed squid and squid-containing products. They also have potential in on-site detection for the advantages of high specificity, high sensitivity, time saving, and ease in operation.
Keywords Squid . Loop-mediated isothermal amplification . Species identification . Rapid detection
Electronic supplementary material The online version of this article (doi:10.1007/s12161-016-0700-6) contains supplementary material, which is available to authorized users. * Junli Feng [email protected]
1
Institute of Seafood, Zhejiang Gongshang University, 149 Jiaogong Road, Hangzhou 310012, China
Introduction As seafood consumption increases, so does public awareness of the associated nutritional and safety issues related to seafood mislabeling (Naaum and Hanner 2015). Food species substitution or mislabeling is one form of fraud which has obvious intention of deceiving the consumers when a lowercost product is labeled as one with a higher value. This practice can also lead to health risks for consumers as different species have varying levels of heavy metals (Lowenstein et al. 2010) and nutritional value (Weaver et al. 2008) or may even be toxic (Cohen et al. 2009). Unfortunately, most consumers are unable to detect cases of mislabeling or fraud, especially when the recognizable external morphological features of fish are removed in the course of processing (Wong and Hanner 2008). On the other hand, with the increasing demand and the globalization of the seafood supply, more fish species are being encountered in the market (Shokralla et al. 2015). Therefore, the need for accurate species identification in seafood has become more critical (Shokralla et al. 2015). Squids, order Teuthoidea, are the most diverse group of cephalopods with over 300 species classified into 28 families (Jereb et al. 2010a, b; Roper and Jereb 2010). According to Food and Agriculture Organization (FAO
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