Rapid simultaneous detection of Salmonella and Campylobacter bacteria directly from chicken faeces

Campylobacter and Salmonella are widely recognized as significant causes of foodborne diseases, with poultry and poultry products being the most frequent sources of infection in humans. Rapid, simultaneous detection of both pathogens can prevent contaminated food from entering the human food chain. The present study aimed to simultaneously detect Salmonella and Campylobacter spp. directly from faeces of broiler chickens aged 4 to 5 weeks by multiplex polymerase chain reaction (mPCR). The suitability of the PCR protocol using pairs of primers targeting the invA and cadF genes was evaluated to detect Salmonella and Campylobacter spp., respectively, from 15 samples. The specificity of the PCR assay was 100% for both pathogens, as no positive cross-reactions were detected with non-targeted bacteria. The limit of detection for pure culture of Salmonella was 1 CFU/mL, whereas for Campylobacter it was 101 CFU/mL. Incubation of spiked faeces in brain heart infusion broth for 24 to 48 hours maintained a detection limit of 1 CFU/mL for Salmonella; however, the sensitivity decreased, resulting in a detection limit of 102 CFU/mL for Campylobacter. Simultaneous detection from 15 chicken faecal samples revealed two samples co-carrying both Salmonella and Campylobacter spp., four samples positive for Salmonella, six samples positive for Campylobacter, and three samples were negative for both pathogens. The developed mPCR protocol in the present study was highly specific and sensitive for detecting Salmonella and Campylobacter spp. directly from chicken faeces, achieving results in under 36 hours compared to the conventional culture method. The mPCR protocol can benefit veterinary and public health authorities during epidemiological investigations and rapid diagnostic purposes, which require timely confirmation of the disease status in chickens during the grow period.

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