O-antigens on the surface of Escherichia coli are important virulence factors that are targets of both the innate and adaptive immune system and play a major role in pathogenicity. O-antigens that are responsible for antigenic specificity of the strain determine the O-serogroup. E. coli O26, O45, O103, O111, O113, O121, O145, and O157 have been the most commonly identified O-serogroups associated with Shiga toxin-producing E. coli (STEC) implicated in outbreaks of human illness all over the world. A multiplex polymerase chain reaction assay was developed to simultaneously detect the eight STEC O-serogroups targeting the wzx (O-antigen-flippase) genes of all O-antigen gene clusters. The sensitivity of the multiplex polymerase chain reaction was found to be 10 colony forming units for each O-group when enriched in broth and 100 colony forming units when enriched in artificially inoculated apple juice diluted with tryptic soy broth for 16 h at 37°C. The method can be used for detecting STEC O-groups simultaneously and may be exploited for improving the safety of food products.
Salmonella enterica is a major food-borne pathogen of world-wide concern. Sensitive and rapid detection methods to assess product safety before retail distribution are highly desirable. Since Salmonella is most commonly associated with poultry products, an evanescent wave fiber-optic assay was developed to detect Salmonella in shell egg and chicken breast and data were compared with a time-resolved fluorescence (TRF) assay. Anti-Salmonella polyclonal antibody was immobilized onto the surface of an optical fiber using biotin-avidin interactions to capture Salmonella. Alexa Fluor 647-conjugated antibody (MAb 2F-11) was used as the reporter. Detection occurred when an evanescent wave from a laser (635 nm) excited the Alexa Fluor and the fluorescence was measured by a laser-spectrofluorometer at 710 nm. The biosensor was specific for Salmonella and the limit of detection was established to be 103 cfu/mL in pure culture and 104 cfu/mL with egg and chicken breast samples when spiked with 102 cfu/mL after 2–6 h of enrichment. The results indicate that the performance of the fiber-optic sensor is comparable to TRF, and can be completed in less than 8 h, providing an alternative to the current detection methods.
Recent outbreaks of food borne illnesses continue to support the need for rapid and sensitive methods for detection of foodborne pathogens. A method for detecting Listeria monocytogenes in food samples was developed using an automated fiber-optic-based immunosensor, RAPTOR ™. Detection of L. monocytogenes in phosphate buffered saline (PBS) was performed to evaluate both static and flow through antibody immobilization methods for capture antibodies in a sandwich assay. Subsequent detection in frankfurter samples was conducted using a flow through immobilization system. A two stage blocking using biotinylated bovine serum albumin (b-BSA) and BSA was effectively employed to reduce the non-specific binding. The sandwich assay using static or flow through mode of antibody immobilization could detect 1×10 3 cfu/ml in PBS. However, the effective disassociation constant K d and the binding valences for static modes of antibody immobilization in spiked PBS samples was 4×10 5 cfu/ml and 4.9 as compared to 7×10 4 cfu/ml and 3.9 for flow through method of antibody immobilization. Thus the sensitive flow-through immobilization method was used to test food samples, which could detect 5×10 5 cfu/ml of L. monocytogenes in frankfurter sample. The responses at the lowest detectable cell numbers in the frankfurter samples was 92.5 ± 14.6 pA for L. monocytogenes to comparative responses of 27.9 ± 12.2 and 31 ± 14.04 pA obtained from Enterococcus Sensors 2006, 6 809 faecalis and Lactobacillus rhamnosus (control species), respectively. The effective K d and binding valency from spiked frankfurter samples was 4.8×10 5 cfu/ml and 3.1, thus showing highly sensitive detection can be achieved using the RAPTOR ™ biosensor even in the presence of other bacterial species in the matrix.
Rapid and sensitive detection tools are essential in pathogen control and prevention in food and in food processing facilities. The conventional culture methods provide reliable results but the analysis time, labor and the cost associated with these methods often pose a great inconvenience to industrial applications. Biosensor‐based detection methods are considered to be promising emerging technologies which are capable of fulfilling the current needs in providing results rapidly with improved sensitivity and specificity. In this review we described three cutting‐edge optical sensor technologies: light scattering, fiber optic‐ and cell‐based sensors, which are proven to be useful in pathogen detection from food samples.
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