A Double Antibody Sandwich ELISA Assay for the Detection of Salmonella in Food Original Samples
نویسندگان
چکیده
Development of monoclonal antibody based sandwich ELISA for rapid detection of Salmonella and optimization of enrichment procedures to increase the sensitivity. Spleen cells from Balb/c mice immunized with flagellin (H=d) antigen of S. typhi were fused with NSO myeloma clones for preparation of mAbs and New Zealand rabbits were used to produce polyclonal antibody. Different enrichment broths were also evaluated. Out of six specific clones to H=d antigen, mAb L2D3G6 was chosen to establish the sELISA curve, which had the detection limit of 104-105 CFU of S. typhi, and the incubation time could be shorten to 4 h. Of the various broths, BPW was found to yield maximum ELISA values. When tested in artificially inoculated food samples, this assay could detect 102 S. typhi CFU/mL within 10 h from various food rinses (meat, vegetable) and milk samples. In comparison to culture, the enrichment-sELISA is rapid and sensitive, and may be used as a rapid screening procedure for environmental monitoring during outbreak situation. KeywordsSalmonella; S. typhi; Monoclonal antibody; Sandwich ELISA
منابع مشابه
Double Sandwich ELISA Modified Method for the Detection of Clostridium Botulinum Type E
Background & Objective: A very small amount of botulinum toxin can cause death and on the other hand, there is no cure for its poison other than antitoxin. Therefore, a diagnostic method that can detect very small amounts of botulinum toxin in a short time is very important. In this study, rapid and accurate detection of botulinum toxin type E has been performed with the double sandwich ELISA m...
متن کاملDevelopment of a double-antibody sandwich ELISA for rapid detection of Bacillus Cereus in food
Bacillus cereus is increasingly recognized as one of the major causes of food poisoning in the industrialized world. In this paper, we describe a sensitive double-antibody sandwich enzyme-linked immunosorbent assay (ELISA) that was developed for rapid detection of B. cereus in food to minimize the risk of contamination. The polyclonal antibody (pAb) and monoclonal antibodies (mAbs) specific to ...
متن کاملDevelopment of a Sandwich ELISA for EHEC O157:H7 Intimin γ1
Enterohemorrhagic Escherichia coli (EHEC) O157:H7 is a zoonotic pathogen of worldwide importance that causes foodborne infections in humans. Intimin gamma 1 (intimin γ1) is one of the most important outer membrane proteins required for EHEC's intimate adhesion to epithelial cells. Here, we generated a polyclonal antibody (pAb) and a monoclonal antibody (mAb) against intimin γ1 to develop a doub...
متن کاملEnzyme-linked immunosorbent assay for detection of staphylococcal enterotoxins in foods.
An enzyme-linked immunosorbent assay (ELISA) was developed for detection of staphylococcal enterotoxins in foods. The "double-antibody sandwich" protocol combines parts of several procedures reported previously. Horseradish peroxidase was conjugated to antibody specific for an enterotoxin, and the antibody-enzyme conjugate was assayed with a 2,2'-azino-di-(3-ethylbenzthiazoline sulfonic acid)-H...
متن کاملPotential Use of DNA Aptamer-Magnetic Bead Separation-PCR Assay for Salmonella Detection in Food
Background: Salmonella is one of the most common food-borne pathogens that can cause illness. In this study, the sensitivity and the specificity of Aptamer-Magnetic bead Separation-Polymerase Chain Reaction (AMS-PCR) method were determined for Salmonella spp. detection. Methods: Different concentrations of Salmonella enterica were mixed with streptavidin-magnetic beads coated with biotinylated...
متن کامل