Application of Loop-Mediated Isothermal Amplification (LAMP) for Rapid Detection of Jumbo Flying Squid Dosidicus gigas (D'Orbigny, 1835)

作者：Ye, Jian; Feng, Junli^*; Dai, Zhiyuan; Meng, Lu; Zhang, Yanping; Jiang, Xiaona

来源：Food Analytical Methods, 2017, 10(5): 1452-1459.

DOI：10.1007/s12161-016-0700-6

• Squid
• Loop-mediated isothermal amplification
• Species identification
• Rapid detection

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 A degrees C for 30 min, only D. gigas DNA can be detected by real-time fluorescence detection (RealAmp) or visual detection via SYBR GreenA (R) 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.

• 出版日期2017-5
• 单位浙江工商大学