A multiplex PCR assay combined with capillary electrophoresis for the simultaneous detection of tropomyosin allergens from oyster, mussel, abalone, and clam mollusk species

Abstract

Tropomyosin present in mollusk species is the most common allergen in humans and causes excessive immune responses. To simultaneously detect tropomyosin allergens in mollusk species, a multiplex PCR assay combined with capillary electrophoresis was developed for the detection of tropomyosin genes of oyster, mussel, abalone, and clam, and the 18S rRNA gene of eukaryotes. The developed multiplex PCR revealed specific amplicons of four mollusk species [oyster (Crassostrea gigas), 150 bp; mussel (Mytilus edulis), 119 bp; abalone (Haliotis discus hannai), 98 bp; clam (Ruditapes philippinarum), 76 bp] and an amplicon of universal eukaryotic primer (eukaryotes, 190 bp); the detection limit of DNA was confirmed to be 16 pg. This multiplex PCR assay was applied for monitoring commercially processed seafood products, achieving successful detection of tropomyosin genes in 19 processed seafood products in Korea. The developed assay is an efficient and useful method for detecting tropomyosin allergens from mollusk species in seafoods.

Keywords: Allergen; Mollusk species; Multiplex PCR; Simultaneous detection; Tropomyosin.