Grimes T M, King D J, Kleven S H
Avian Dis. 1976 Apr-Jun;20(2):299-301.
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.
开发了一种微量滴定细胞培养方法，并用于滴定病毒分离株以进行鉴定。将病毒稀释液和鸡肾细胞悬液加入一次性微量培养板的孔中，在第5天或第6天通过显微镜确定感染终点，或在第6天读取结晶紫染色的单层细胞来确定感染终点。用这种方法，从诊断样本中分离出的37株候选禽腺病毒被鉴定为禽腺病毒（AAV）。用于鉴定的标准包括产生圆形细胞病变效应、对氯仿处理有抗性、被5-溴脱氧尿苷抑制，以及通过琼脂凝胶沉淀存在与已知AAV具有同一性的抗原。对三种AAV的八次重复滴定进行统计分析表明，该滴定方法具有高度可重复性。使用微量培养方法进行滴定可在指示细胞、培养基、培养箱空间、培养皿和时间方面大幅节省。
