Abstract:
Introduction: Increased demand for seafood and limited fishing capacity of wild fish has led to the development of fish farming, but the excessive import of eggs, breeders and ornamental fish has led to the spread of viral diseases in the country and several cases of viral losses and damage to aquaculture have been reported. VHS is one of the notifiable viral diseases of cold-water fish that has spread in recent years.
Objective: Because common diagnostic methods such as cell culture and molecular methods are time consuming and costly so rapid diagnosis of viral diseases is a suitable method for disease controlling, this project was designed to obtain technical knowledge of producing and evaluating hemorrhagic septicemia diagnosis kit.
Methods: Different combination forms of commercial monoclonal antibody and polyclonal antibody produced in New Zealand rabbits was employed for designing the chromatographic test.
Results: In this study, the optimal concentration used for conjugation of gold nanoparticles was 5 μg / ml. Similarly, 1.8 mg / ml polyclonal antibody was used for spraying on the nitrocellulose membrane test line after optimization. For the reaction time in this study, the times of 15, 20 and 25 minutes were compared and it was found that the sensitivity was similar between 20 and 25, but more than 15 minutes. Therefore, the reaction time of this immunochromatographic strip was set at 20 minutes.
Conclusion: Cell culture and molecular methods are commonly used to detect viruses, which are time consuming and require to laboratory equipment and well trained technician. Therefore, to reduce the costs of the screening program, using the designed rapid diagnostic methods is introduced as a quick method for VHSV diagnosis.
Keywords: VHSV, Antibody, Gold nanoparticles, Rapid diagnostic kit