Development of Molecular Detection Kit for Larimichthys crocea and Larimichthys polyactis

Abstract

Large yellow croaker (Larimichthys crocea) and redlip yellow croaker (L. polyactis) are commonly consumed species in fisheries markets in East Asian countries. They have wide differences of price due to consumer preferences for L. polyactis. However, it is difficult to identify the species based on the appearance of both species from the general public. In this study, a real-time polymerase chain reaction (PCR) kit was designed to distinguish these two species. Species-common primer pairs were designed, and two species-specific probes were designed with sequences that were 2 bp different. The L. crocea target probe was modified using HEX dye, whereas the L. polyactis was modified using FAM dye. These primer pairs and probes worked in a single reaction. Based on the confirmation of a simple peak image, both species were sufficiently distinguished. The PCR kit developed in our study can be suggested as a good tool to distinguish both fish species for consumer as well as producer. In addition, the species-specific probes can be used as the biomarker, which applies to the lab-on-a-chip technology