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Nurhidayah and Ince Ayu Khaerana Kadriah
Research Institute for Coastal Aquaculture
Jl. Makmur Dg. Sitakka No. 129, Maros, South Sulawesi,
Indonesia,  90512

This study was aimed to analyze the optimum condition of ELISA method as a diagnostic tool using monoclonal antibody Vibrio harveyi. This study was conducted in the Laboratory of Fish Health and Environment belongs to The Research Institute for Coastal Aquaculture. Temperature of micro plate and a series of different concentrations of monoclonal antibody, as well as concentrations of conjugated were applied as the treatments. The incubation of coating plate were conducted under 4oC and room temperature, whereas the concentrations of monoclonal antibody were 1:25, 1:50 and 1:100; concentrations of conjugated were 1:500, 1:1000, 1:2000 and 1:4000. All treatments were replied for four replications.

The results of this study showed that the values of optic density of each treatment observed using A450 nm were different. The concentrations of monoclonal antibody of 1:25 at 4oC of coating plate were lower compared to concentrations 1:50 and 1:100 at all conjugated (Figure 1). In contrast, higher concentrations of monoclonal antibody were observed at warmer coating temperature (Figure 2). Incubation temperature of 4oC showed better results compared to that observed on room temperature. The monoclonal antibody produced in this study was able to diagnose V. harveyi through ELISA method using optimal concentrations of monoclonal antibody 1:50 elaborated with 1:500 of conjugated under 4oC of coated plate.

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