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

Bergmann, Sven GND; Engler, C.; Wang, Q.; Zeng, W.; Li, Y.; Wang, Y.; Lee, P. Y.; Lindenberger, C.; Reichert, M.; Matras, M.; Fuchs, Walter GND; Reiche, Sven GND; Dauber, Malte GND; Lenk, Matthias GND; Morin, T.; Klafack, S.; Jin, Y.; Monaghan, S.; Kempter, J.

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.

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Bergmann, Sven / Engler, C. / Wang, Q. / et al: Investigation on Antigen ELISA for Detection of the Envelope Glycoprotein Coded By ORF 149 of Different Koi Herpesvirus Isolates Obtained From Cell Cultures. 2017.

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