Application of a microtiter cell-culture method to characterization of avian adenoviruses

A microtiter cell-culture method was developed and used to titrate virus isolates for characterization. Virus dilutions and chicken kidney cell suspensions were dispensed into the wells of disposable microculture plates, with infectivity endpoints being determined microscopically on the fifth or sixth day, or by reading crystal-violet-stained monolayers on day 6. With this method, 37 candidate avian adenoviruses isolated from diagnostic accessions were characterized as avian adenoviruses (AAV). The criteria used for characterization were production of round-cell cytopathic effect, resistance to chloroform treatment, inhibition by 5-bromodeoxyuridine, and the presence of an antigen showing identity with a known AAV by precipitation in agar gel. Statistical anlaysis of eight replicate titrations of three AAV indicated that the titration method was highly reproducible. Use of the microculture method for titrations gave substantial savings in indicator cells, media, incubator space, culture dishes, and time.