Kauffman P E, Shah D B
Food and Drug Administration, Division of Microbiology, Cincinnati, OH 45226.
J Assoc Off Anal Chem. 1988 May-Jun;71(3):636-42.
An enzyme immunoassay (EIA) was developed to identify and quantitate soluble antigens originating from Drosophila melanogaster eggs. Polystyrene microtiter plates were coated with anti-egg antibody. Egg antigen standards or samples were reacted with the sensitized wells. The immobilized antigen was reacted with biotinylated antibody, and the complex was quantitated by an avidin-horseradish peroxidase conjugate. The assay can detect egg antigens equivalent to 0.03 eggs/mL. The antibody was directed against a number of antigens and cross-reacted with extracts of adult Drosophila melanogaster flies. Homogenized eggs diluted in various juice samples were detectable by EIA. Heating the antigen diluted in buffer or juice at a temperature used in processing of juice reduced the immunological response approximately 40-45%. Brine separated from fresh-packed sauerkraut showed EIA responses equivalent to 0.3-3 eggs/mL. Electrophoretic, chromatographic, and immunological analyses suggested that the EIA responses for sauerkraut brine were related to the Drosophila melanogaster egg antigens.
开发了一种酶免疫测定法(EIA)来鉴定和定量源自黑腹果蝇卵的可溶性抗原。聚苯乙烯微量滴定板用抗卵抗体包被。卵抗原标准品或样品与致敏孔反应。固定化抗原与生物素化抗体反应,复合物用抗生物素蛋白-辣根过氧化物酶偶联物进行定量。该测定法可检测相当于0.03个卵/毫升的卵抗原。该抗体针对多种抗原,并与黑腹果蝇成虫提取物发生交叉反应。用EIA可检测稀释在各种果汁样品中的匀浆卵。在果汁加工所用温度下加热缓冲液或果汁中稀释的抗原,免疫反应降低约40-45%。从新鲜包装的酸菜中分离出的盐水显示EIA反应相当于0.3-3个卵/毫升。电泳、色谱和免疫分析表明,酸菜盐水的EIA反应与黑腹果蝇卵抗原有关。