Multiplex PCR (mPCR) for the Detection of Salmonella spp. and the Differentiation of the Typhimurium and Enteritidis Serovars in Chicken Meat

Abstract

Salmonella enterica subspecies enterica serovars Enteritidis and Typhimurium are important causes of foodborne illness. Methods for simultaneous detection of these serovars may contribute for the adoption of measures to prevent these diseases. The polymerase chain reaction to detect individually or simultaneously the serovars Enteritidis and Typhimurium in foods have been standardized; however, the majority of assays employ the fliC gene as a target for detection of serovar Typhimurium. The detection of these sorovars in a few hours allows the food supply chain to take appropriate measures to prevent the distribution of contaminated food. The aim of this study was to develop a new multiplex PCR (mPCR) for the simultaneous detection and differentiation of Salmonella spp., S. Enteritidis and S. Typhimurium in chicken meat. The mPCR assays showed high specificity and differentiated S. Typhimurium from 22 Salmonella serovars tested, including S. Kentucky, showing that it's an alternative to reduce the time required to obtain presumptive positive results. PRACTICAL APPLICATIONSThe fliC gene used as target for the detection of serovar Typhimurium is questionable as it has also been described for S. Kentucky. The amplification of STM4492 gene in this study, instead of fliC gene, exhibited high specificity to detect S. Typhimurium. The developed mPCR is an efficient means for the simultaneous detection and differentiation of Salmonella spp., S. Enteritidis and S. Typhimurium in chicken meat after 24 h of enrichment. The mPCR has the potential to be used in routine diagnostic laboratories to obtain presumptive positive results and for identification of Enteritidis and Typhimurium strains isolated by the conventional method. bs_bs_banner Journal of Food Safety ISSN 1745-4565

Multiplex-PCR Assay for Detection of some Major Virulence Genes of <i>Salmonella enterica</i> Serovars from Diverse Sources

“…PCR assays have been developed for the detection of different gene-encoded virulence factors, viz. simplex PCR for Salmonella enterotoxin (stn) 12 , Salmonella enteritidis fimbriae (sef ) and plasmid encoded fimbriae (pef ) 13 , and multiplex PCR (m-PCR) assays for a combination of a few such virulence factors [14][15][16] . Here, we report the development of a m-PCR for rapid and simultaneous detection of seven major virulence genes of S. enterica.…”

Salmonella Enteritidis predominance determined by serotyping andreal-time PCR in poultry-derived food and avian isolates

“…A recent study from Turkey by Dumen et al (12) also reported SE as the most prevalent (26.6%) serotype both by PCR and by conventional serotyping. In contrast, this serovar was either not detected (7,8) or was detected in very low rates (6) from chicken meat-related or egg samples by conventional or real-time PCR in different studies.…”

“…Differences in the detection rate of SE by rPCR in this study compared to the findings of others could mainly be related to the country and prevalence of the serovar in that region during that time period. In addition, sample size and type, and the validity of the PCR detection system (specificity, sensitivity, accuracy/reliability), can be the main factors among others affecting this outcome (7,8,30).…”

“…This useful but time-consuming and labor-intensive method, despite its inherent disadvantages, is commonly used in initial screening of the isolate, followed by molecular subtyping for strain identification (4,5). Therefore, for rapid detection of SE and other frequently reported Salmonella serotypes, alternative serotype-specific conventional or real-time PCRs (rPCRs) were performed and reported (6)(7)(8)(9) to augment taking immediate actions related to public health.…”

Detection of viable hookworm ova from wastewater and sludge

“…Multiplex PCR (mPCR) method can be used to detect multiple parasites including different species of hookworm in a single assay ( Verweij et al, 2007;Jonker et al, 2012;Saeki et al, 2013) and reduce detection time, costs and labor that is otherwise associated with running multiple assays (Toplak et al, 2012). However, it should be noted that multiple primers pairs must function under the same reaction conditions and do not form primer dimers during the reaction.…”