title: Identification of four squid species by quantitative real-time polymerase chain reaction.
abstract: Squids are distributed worldwide, including many species of commercial importance, and they are often made into varieties of flavor foods. The rapid identification methods for squid species especially their processed products, however, have not been well developed. In this study, quantitative real-time PCR (qPCR) systems based on specific primers and TaqMan probes have been established for rapid and accurate identification of four common squid species (Ommastrephes bartramii, Dosidicus gigas, Illex argentinus, Todarodes pacificus) in Chinese domestic market. After analyzing mitochondrial genes reported in GenBank, the mitochondrial cytochrome b (Cytb) gene was selected for O. bartramii detection, cytochrome c oxidase subunit I (COI) gene for D. gigas and T. Pacificus detection, ATPase subunit 6 (ATPase 6) gene for I. Argentinus detection, and 12S ribosomal RNA (12S rDNA) gene for designing Ommastrephidae-specific primers and probe. As a result, all the TaqMan systems are of good performance, and efficiency of each reaction was calculated by making standard curves. This method could detect target species either in single or mixed squid specimen, and it was applied to identify 12 squid processed products successfully. Thus, it would play an important role in fulfilling labeling regulations and squid fishery control.
authors.affiliation: Institute of Seafood, Zhejiang Gongshang University, Hangzhou, Zhejiang 310012, PR China.
authors.forename: Jian
authors.initials: J
authors.lastname: Ye
authors.affiliation: Institute of Seafood, Zhejiang Gongshang University, Hangzhou, Zhejiang 310012, PR China.
authors.forename: Junli
authors.initials: J
authors.lastname: Feng
authors.affiliation: Institute of Bioengineering, Zhejiang Sci-Tech University, Hangzhou, Zhejiang 310018, PR China.
authors.forename: Shasha
authors.initials: S
authors.lastname: Liu
authors.affiliation: Institute of Seafood, Zhejiang Gongshang University, Hangzhou, Zhejiang 310012, PR China.
authors.forename: Yanping
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Institute of Seafood, Zhejiang Gongshang University, Hangzhou, Zhejiang 310012, PR China.
authors.forename: Xiaona
authors.initials: X
authors.lastname: Jiang
authors.affiliation: Institute of Seafood, Zhejiang Gongshang University, Hangzhou, Zhejiang 310012, PR China. Electronic address: dzy@zjgsu.edu.cn.
authors.forename: Zhiyuan
authors.initials: Z
authors.lastname: Dai
mesh_terms: D000818:Animals; D045303:Cytochromes b / Q000235:genetics; D017931:DNA Primers; D004272:DNA, Mitochondrial / Q000737:chemistry / Q000235:genetics*; D049832:Decapodiformes / Q000145:classification / Q000235:genetics*; D003576:Electron Transport Complex IV / Q000235:genetics; D025261:Mitochondrial Proton-Translocating ATPases / Q000235:genetics; D012335:RNA, Ribosomal / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D017422:Sequence Analysis, DNA / Q000379:methods*; D013045:Species Specificity
chemical_list: D017931:DNA Primers; D004272:DNA, Mitochondrial; D012335:RNA, Ribosomal; C075526:RNA, ribosomal, 12S; D045303:Cytochromes b; D003576:Electron Transport Complex IV; D025261:Mitochondrial Proton-Translocating ATPases
keywords: Mitochondrial DNA; Quantitative real-time PCR; Species identification; Squid