Sensitivity of enrichment-PCR method for Salmonella enterica serovar Typhimurium and Salmonella enterica serovar Enteritidis analysis in chicken carcasses

Nurjanah S., . and Wulan H.A., . and Rahayu W. P., . Sensitivity of enrichment-PCR method for Salmonella enterica serovar Typhimurium and Salmonella enterica serovar Enteritidis analysis in chicken carcasses. pp. 54-61. ISSN 2550-2166

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.

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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.

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Item Type:	Article
AGROVOC Term:	Chickens
AGROVOC Term:	Salmonella
AGROVOC Term:	Pathogenic bacteria
AGROVOC Term:	Food enrichment
AGROVOC Term:	Salmonella typhimurium
AGROVOC Term:	Salmonella enteritidis
AGROVOC Term:	PCR
AGROVOC Term:	Carcases
AGROVOC Term:	Food sensitivity
AGROVOC Term:	Gram negative bacteria
Depositing User:	Mr. AFANDI ABDUL MALEK
Last Modified:	24 Apr 2025 00:55
URI:	http://webagris.upm.edu.my/id/eprint/10388

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