Abstract
Salmonella spp. is Gram negative-pathogenic bacteria that usually found as a contaminant in chicken carcasses. This study was aimed to increase the sensitivity of PCR enrichment step and apply the enrichment-PCR combination to detect Salmonella in chicken carcasses. In this study were used Salmonella enterica serovar Hadar, Salmonella enterica serovar Typhimurium and Salmonella enterica serovar Enteritidis with the target genes were invA, STM4497, and respectively. A total of 25 g of the chicken carcasses were artificially contaminated by approximately 0.96 and 3.33 MPN/mL for each serovar separately. Samples were incubated in pre-enrichment and enrichment media for 8 hrs prior to the DNA extraction. The pre-enrichment and enrichment media was Buffered Peptone Water and Rappaport-Vassiliadis-soya. The result showed that the target genes of S. enterica ser. Hadar, S. enterica ser. Typhimurium and S. enterica ser. Enteritidis were detected in chicken carcasses, indicated by the presence of DNA band with the size was 429 bp, 311 bp and 135 bp respectively. These result in line with analysis using ISO method and BLAST-comparison analysis of DNA amplicon sequences with GenBank references. Application of this method for Salmonella detection in chicken carcasses sold in the traditional market showed a higher prevalence than the previous result without enrichment. All samples (n = 100) from unsanitary practice sellers were positively contaminated by Salmonella spp. and also high prevalence for S. enterica ser. Typhimurium and S. enterica ser. Enteritidis. It can be concluded that enrichment is an important step to increase the sensitivity detection of PCR method.
Highlights
Salmonella spp. is facultative anaerobic bacteria, Gram-negative, and rod-shaped bacilli
This study aimed to increase the sensitivity of Salmonella spp. detection
The incubation time of 2 hrs was not enough to detect these bacteria at a low number
Summary
Salmonella spp. is facultative anaerobic bacteria, Gram-negative, and rod-shaped bacilli. This bacterium belongs to family Enterobacteriaceae (Kim et al, 2006). More than 2600 serovars Salmonella spp. have been identified (Popoff et al, 2003). Salmonella has been known as a foodborne pathogen and caused outbreaks in several countries. Salmonella infections occurred in America (Scallan et al, 2011), China (Xiao et al, 2015), Australia (Ford et al, 2018), Canada (Morton et al, 2019). Salmonella enterica serovar Typhimurium and Salmonella enterica serovar Enteritidis are the dominant contaminant serovars in chicken carcasses. Both serovars are reported being the main cause of salmonellosis in humans (Lee et al, 2009)
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