Table 1.
Bacterial strains used in this study.
Figure 1.
Sandwich-type format carried out in ELISA plate wells for the capture and detection of Vibrio cells.
The well surface is coated with neutravidin and blocked with bovine serum albumine. The biotinylated anti-vibrio polyclonal antibody is added and bacterial cells are then captured and detected using an horse radish peroxidase linked anti-vibrio polyclonal antibody that can be detected colorimetrically using a spectrophotometer at 450 nm.
Figure 2.
Antibody affinity for different Vibrio strains.
Absorbance signals obtained after direct adsorption of bacterial strains to the well surface for one hour followed by detection using a 1/500 dilution of horseradish peroxidase anti-Vibrio antibody (HRP-αVib Pab) incubated for one hour onto the surface.
Table 2.
LOD, IC50 and sensitivity values obtained for each Vibrio strain with or without functionalising the plate surface.
Figure 3.
Affinity of the different Vibrio strains and commercial positive control to the functionalised surface using optimised conditions.
Absorbance signals obtained after a 30 min cell capture step on the functionalised surface and a 30 min detection step using a 1/1000 dilution of horseradish peroxidase anti-Vibrio antibody (HRP-αVib Pab).
Figure 4.
Signals obtained for increasing Vibrio parahaemolyticus (dashed line) or S. marcescens (solid line) cell concentrations when carrying out the capture step either in PBS (▪), filtered (▴) or unfiltered (•) natural seawater.
Figure 5.
Linear part of the ELISA standard curve.
Absorbance values obtained for increasing V. parahaemolyticus cell concentrations ranging from 1×104 to 1×108 cells ml−1.
Table 3.
Comparison of the Vibrio counts obtained using both the TCBS plate culturing and ELISA detection method.