A PCR- ELISA for the detection of Salmonella from chicken intestine

Abstract

In this study, a Polymerase Chain Reaction Enzyme Linked Immunosorbent Assay (PCR-ELISA) was described to detect Salmonella DNA from selective primary enrichment culture of chicken intestine. Salmonella genus-specific PCR product was produced using invA-specific primers and digoxigenin by commercial PCR ELISA DIG Labelling and ELISA DIG Detection kits. PCR-ELISA detection limits with S. Enteritidis 64K DNA, pure culture, and with the intestinal homogenate artificiallycontaminated with this strain were found as 0.0079 µg ml-1, 50 cfu ml-1, and 70 cfu ml-1, respectively. Sensitivity and specificity of the assay was determined as 100% with the tests performed with 41 Salmonella enterica serovar DNAs, with non-Salmonella strains, Citrobacter sp., E. coli, Klebsiella sp., P. aeruginosa, and Streptococcus sp. DNAs, and with 124 tetrathionate broth enrichment cultures of Salmonella-contaminated chicken intestinal samples. We suggest that this PCR-ELISA detection could be an alternative method to detect Salmonella-specific DNA from chicken intestine, and it would find use particularly in highsample throughput laboratories in the poultry sector.