Use of Genetic and Immunological Probes for Pediocin PA-1 Gene Detection and Quantification of Bacteriocin Production in Pediococcus acidilactici Strains of Meat Origin

Abstract

PCR amplification analysis with pediocin PA-1 (pedA1) specific primers, a gene probe covering the pedA1 structural gene, and an immunological probe based on anti-peptide antibodies with predetermined specificity for pedA1, have been evaluated for pedA1 structural gene detection and quantification of bacteriocin production in Pediococcus acidilactici strains of meat origin. PCR products were obtained from colonies and purified plasmid DNA. The gene probe was used in dot-blot and colony-blot hybridization analysis. The production of pedA1 was evaluated by a non-competitive indirect ELISA (NCI-ELISA), a competive indirect ELISA (CI-ELISA), a competitive direct ELISA (CD-ELISA), and by immunoblotting. The approaches taken in the generation and performance of the reported probes could be useful for the rapid detection and identification of other bacteriocin-producing lactic acid bacteria (LAB) from many substrates and reservoirs and for quantification of bacteriocin production in food and environmental isolates and heterologous hosts.