Investigation on Antigen ELISA for Detection of the Envelope Glycoprotein Coded By ORF 149 of Different Koi Herpesvirus Isolates Obtained From Cell Cultures

An antigen capture ELISA was established for cell culture materials using a monoclonal (mab) antibody 10A9 which recognized the product of KHV ORF 149. Five KHV isolates were replicated onto CCB cell cultures in six-well plates for eight days at 20 and at 26 °C incubation temperature. In the beginning, samples were taken from the different isolates at 1, 2, 4, and 6 hours post infection, afterwards one plate was collected per day. All samples were investigated by different PCRs and RT-PCR and by antigen ELISA. The results were compared. While the isolates replicated at 20 °C developed a two cycle replication at 1 dpi and a second between 4 and 5 dpi, the same isolates replicated at 26 °C developed only a single peak between 5 and 6 dpi. This could be shown on DNA level by qPCR and on protein/antigen level by ELISA, but not on RNA level.