The development of a hexaplex-conventional PCR for identification of six animal and plant species in foodstuffs

Abstract

A hexaplex-conventional PCR assay was developed for identification of five meat and one plant species origins in foodstuffs simultaneously. The method merges the use of horse (Equus caballus), soybean (Glycine max), sheep (Ovis aries), poultry (Meleagris meleagris), pork (Sus scrofa), and cow (Bos taurus) specific primers that amplify fragments (horse; 85 bp, soybean; 100 bp, sheep; 119 bp, poultry; 183 bp, pork; 212 bp and cow; 271 bp) of the mitochondrial cyt b, lectin, 12S rRNA, 12S rRNA, ATPase subunit 6 genes and ATPase subunit 8 genes respectively, and a universal 18S rRNA primers that amplifies a 141 bp. Multiplex analysis of the reference food samples showed that detection limit of the hexaplex assay was 0.01% for each species. Taken together, all data indicated that this hexaplex PCR assay was a simple, fast, sensitive, specific, and cost-effective detection method for horse, soybean, sheep, poultry, pork and cow species in foodstuffs.

Keywords: 18S rRNA; Foodstuffs; Hexaplex-conventional PCR; Mitochondrial DNA; Species origins identification; Species-specific primers.