Development of a Multiplex PCR System for the Simultaneous Detection of the Shrimp Species Fenneropenaeus chinensis, Litopenaeus vannamei, and Penaeus monodon

Due to mixed use of shrimp in foodstuffs, it is important to distinguish Fenneropenaeus chinensis from other shrimp species, such as Litopenaeus vannamei and Penaeus monodon. For this purpose, we have developed a multiplex PCR method to detect the simultaneous presence of all three species. Universal primers specific to the 16S ribosomal RNA subunit were used to sequence and ascertain the species in the samples. In this study, specific primers were designed to result in a unique PCR product size (143, 260, and 348 bp for L. vannamei, F. chinensis and P. monodon, respectively). In addition, duplex and triplex PCR protocols were developed to concurrently and rapidly detect these three shrimp species without nonspecific gene amplification. This multiplex PCR system is expected to be widely used to distinguish shrimp species that should be monitored to prevent their mixed use in food.