Implementation of real-time PCR to tetrathionate broth enrichment step of Salmonella detection in poultry

Abstract

Aims: The present study describes the implementation of real-time PCR to tetrathionate broth enrichment step of Salmonella detection in poultry.

Methods and Results: Real-time PCR with Salmonella invA-specific primers and a standard bacteriological method was applied to detect Salmonella in tetrathionate enrichment cultures of 492 intestinal homogenates and 27 drag swabs from 47 poultry flocks. The number of positive individual samples by real-time PCR and culture method was 65 (12.5%) and 35 (6.8%), respectively. The number of Salmonella-positive flocks was 13 (27.7%) by both methods. PCR detection required 25 min for up to 32 samples. Melting curve analysis revealed the T, for Salmonella-specific PCR product as 87 +/- 1degreesC.

Conclusions: Implementation of real-time PCR to tetrathionate broth enrichment step reduces the Salmonella detection time to 18 h and 25 min. Isolation of Salmonella should be carried out with PCR to determine the serovar.

Significance and Impact of the Study: Real-time PCR is a powerful tool in rapid and accurate Salmonella monitoring in poultry companies, together with standard bacteriology.