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Salmonella is one of major foodborne pathogens and the leading cause of foodborne illness-related hospitalizations and deaths. It is critical to develop a sensitive and rapid detection assay that can identify Salmonella to ensure food safety. In this study, a DNA sensor-based suspension array system of high multiplexing ability was developed to identify eight Salmonella serovars commonly associated with foodborne outbreaks to the serotype level. Each DNA sensor was prepared by activating pre-encoded microspheres with oligonucleotide probes that are targeting virulence genes and serovar-specific regions. The mixture of 12 different types of DNA sensors were loaded into a 96-well microplate and used as a 12-plex DNA sensor array platform. DNA isolated from Salmonella was amplified by multiplex polymerase chain reaction (mPCR), and the presence of Salmonella was determined by reading fluorescent signals from hybridization between probes on DNA sensors and fluorescently labeled target DNA using the Bio-Plex® system. The developed multiplex array was able to detect synthetic DNA at the concentration as low as 100 fM and various Salmonella serovars as low as 100 CFU/mL within 1 h post-PCR. Sensitivity of this assay was further improved to 1 CFU/mL with 6-h enrichment. The array system also correctly and specifically identified serotype of tested Salmonella strains without any cross-reactivity with other common foodborne pathogens. Our results indicate the developed DNA sensor suspension array can be a rapid and reliable high-throughput method for simultaneous detection and molecular identification of common Salmonella serotypes.

In this study, the presence of extended spectrum β-lactamase (ESBL)-producing Escherichia coli in aquatic environments (the Orontes River and an urban wastewater) was investigated. Fifty-four E. coli strains resistant to cefotaxime were isolated from the river waters and nearby waste water treatment plant and screened for ESBL gene variants, different classes of integrons and sulfonamide resistance genes. The ESBL-producing E. coli strains were further characterized by PhP-typing system, phylogenetic grouping and antimicrobial susceptibility testing. Of the 54 ESBL-producing strains, 14 (25.9%) belonged to four common PhP types and the remaining were of single types. CTX-M type ESBL genes were identified in 68% of the isolates. The most predominant specific CTX-M subtype identified was blaCTX−M−15 (n = 36), followed by blaCTX−M−1 (n = 1). None of the isolates were SHV and OXA positive. Most of the ESBL positive isolates (n = 37; 68.5%) were harboring sul gene. This study indicates a widespread distribution of CTX-M-15 producing E. coli strains in the surface waters in part of Turkey, suggesting an aquatic reservoir for ESBL genes.

Salmonella is the leading cause of bacteria-associated foodborne illnesses in the United States. Early detection of this pathogen by a rapid and sensitive assay is important to prevent salmonellosis. In this study, we describe a magnetic bead-based immunoassay for detection of Salmonella consisting of immunomagnetic separation for simple target concentration with tyramide signal amplification to increase the assay sensitivity. The developed immunoassay was able to detect Salmonella Typhimurium in culture with the detection limit of 280 CFU/mL in less than 3 h without any enrichment and further decreased to 70 CFU/mL with 3 h enrichment. When tested with ground beef and poultry samples artificially contaminated with S. Typhimurium and Enteritidis, the assay showed increased detection limits with 800 and 200 CFU/mL, respectively, due to the effect of complex food matrices. However, when 12 h enrichment was added, the detection limits in both food matrices decreased to 1 CFU. Practical Applications This study demonstrated that using immunomagnetic separation (IMS) and tyramide signal amplification can enhance sensitivity and specificity for the detection of Salmonella in food samples. The developed assay has great potential as a simple monitoring system for foodborne pathogens in food samples, which can improve food safety and public health. The results were also compared with sandwich type enzyme-linked immunosorbent assay, which showed the developed assay is approximately 50 times more sensitive than enzyme-linked immunosorbent assay.

Salmonella is the leading cause of foodborne illnesses in the United States, and one of the main contributors to salmonellosis is the consumption of contaminated poultry and poultry products. Since deleterious effects of Salmonella on public health and the economy continue to occur, there is an ongoing need to develop more advanced detection methods that can identify Salmonella accurately and rapidly in foods before they reach consumers. Rapid detection and identification methods for Salmonella are considered to be an important component of strategies designed to prevent poultry and poultry product-associated illnesses. In the past three decades, there have been increasing efforts towards developing and improving rapid pathogen detection and characterization methodologies for application to poultry and poultry products. In this review, we discuss molecular methods for detection, identification and genetic characterization of Salmonella associated with poultry and poultry products. In addition, the advantages and disadvantages of the established and emerging rapid detection and characterization methods are addressed for Salmonella in poultry and poultry products. The methods with potential application to the industry are highlighted in this review.

Escherichia coli O157:H7 is a major foodborne pathogen that has posed serious problems for food safety and public health. Recent outbreaks and recalls associated with various foods contaminated by E. coli O157:H7 clearly indicate its deleterious effect on food safety. A rapid and sensitive detection assay is needed for this harmful organism to prevent foodborne illnesses and control outbreaks in a timely manner. We developed a magnetic bead-based immunoassay for detection of E. coli O157:H7 (the most well-known Shiga toxigenic E. coli strain) with a 96-well microplate as an assay platform. Immunomagnetic separation (IMS) and tyramide signal amplification were coupled to the assay to increase its sensitivity and specificity. This immunoassay was able to detect E. coli O157:H7 in pure culture with a detection limit of 50 CFU/ml in less than 3 h without an enrichment step. The detection limit was decreased 10-fold to 5 CFU/ml with addition of a 3-h enrichment step. When this assay was tested with other nontarget foodborne pathogens and common enteric bacteria, no cross-reactivity was found. When tested with artificially contaminated ground beef and milk samples, the assay sensitivity decreased two- to fivefold, with detection limits of 250 and 100 CFU/ml, respectively, probably because of the food matrix effect. The assay results also were compared with those of a sandwich-type enzyme-linked immunosorbent assay (ELISA) and an ELISA coupled with IMS; the developed assay was 25 times and 4 times more sensitive than the standard ELISA and the IMS-ELISA, respectively. Tyramide signal amplification combined with IMS can improve sensitivity and specificity for detection of E. coli O157:H7. The developed assay could be easily adapted for other foodborne pathogens and will contribute to improved food safety and public health.