Use of genetic and immunological probes for pediocin PA-1 gene detection and quantification of bacteriocin production in Pediococcus acidilactici strains of meat origin
Jm. Martinez et al., Use of genetic and immunological probes for pediocin PA-1 gene detection and quantification of bacteriocin production in Pediococcus acidilactici strains of meat origin, FOOD AGR IM, 12(4), 2000, pp. 299-310
PCR amplification analysis with pediocin PA-1 (pedA1) specific primers, a g
ene probe covering the pedA1 structural gene, and an immunological probe ba
sed on anti-peptide antibodies with predetermined specificity for pedA1, ha
ve 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 gen
e probe was used in dot-blot and colony-blot hybridization analysis. The pr
oduction of pedA1 was evaluated by a non-competitive indirect ELISA (NCI-EL
ISA), 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.