Abstract
BackgroundImproved methods with better separation and concentration ability for detection of foodborne pathogens are in constant need. The aim of this study was to evaluate microplate immunocapture (IC) method for detection of Salmonella Typhi, Shigella flexneri and Vibrio cholerae from food samples to provide a better alternative to conventional culture based methods.ResultsThe IC method was optimized for incubation time, bacterial concentration, and capture efficiency. 6 h incubation and log 6 CFU/ml cell concentration provided optimal results. The method was shown to be highly specific for the pathogens concerned. Capture efficiency (CE) was around 100% of the target pathogens, whereas CE was either zero or very low for non-target pathogens. The IC method also showed better pathogen detection ability at different concentrations of cells from artificially contaminated food samples in comparison with culture based methods. Performance parameter of the method was also comparable (Detection limit- 25 CFU/25 g; sensitivity 100%; specificity-96.8%; Accuracy-96.7%), even better than culture based methods (Detection limit- 125 CFU/25 g; sensitivity 95.9%; specificity-97%; Accuracy-96.2%).ConclusionThe IC method poses to be the potential to be used as a method of choice for detection of foodborne pathogens in routine laboratory practice after proper validation.
Highlights
Improved methods with better separation and concentration ability for detection of foodborne pathogens are in constant need
This study focused on three important foodborne pathogens: Vibrio cholerae, Salmonella Typhi and Shigella flexneri
To analyze the capture efficiency of the microplate IC method, bacterial cell was added to the wells and incubated for varying periods (1-24 h)
Summary
Improved methods with better separation and concentration ability for detection of foodborne pathogens are in constant need. The microbiological analysis of food involves culture enrichment followed by isolation on selective media [2]. The initial pre-enrichment of a food sample allows for resuscitation of physiologically stressed microbes and grows all bacteria to detectable levels (>10 CFU/ml), followed by a period of selective. Fakruddin et al BMC Microbiology (2017) 17:189 in food which cannot be detected by the conventional culture based method [5]. There is a great need for improved methods for foodborne pathogen detection in food matrices. Concentration and separation of pathogens from the food matrix has been the focus of many studies investigating ways to improve sample assay detection limits and speed time to results
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