Molecular identification of the black tiger shrimp ( Penaeus monodon), the white leg shrimp ( Litopenaeus vannamei) and the Indian white shrimp ( Fenneropenaeus indicus) by PCR targeted to the 16S rRNA mtDNA

Abstract

Polymerase chain reaction-based methodologies have been developed for the identification of three commercially-relevant penaeid shrimp species, these were: Litopenaeus vannamei, Penaeus monodon and Fenneropenaeus indicus in food products. Such three species represent more than 80% of the whole farmed shrimp production worldwide and may be fraudulently replaced by species exhibiting lower value such as Litopenaeus stylirostris, Penaeus semisulcatus and Fenneropenaeus merguiensis, respectively. For it, preliminary sequencing of a mitochondrial sequence of ca. 530 bp in the 16S rRNA/tRNA Val mitochondrial region was performed in nearly 20 penaeid shrimp species of commercial relevance. Careful analysis of such sequences allowed the design of primers PNVF/PNVR, which allowed the combined identification of P. monodon and L. vannamei, and PNIF/PNIR, which allowed the specific identification of F. indicus. In addition, P. monodon and L. vannamei could be easily differentiated by either restriction with TspE1 or by amplification with novel primers MPNF/MPNR, specific for P. monodon. The proposed specific methods improve current general identification methods of these species based on more general RFLP analyses. In addition, these methods can be easily completed in less than 8 h.