Blanchard G C, Gardner R E
J Immunol Methods. 1982;52(1):81-90. doi: 10.1016/0022-1759(82)90352-0.
A fluoroimmunoassay (FIA) for the measurement of immunoglobulin E (IgE) is described. The method involves a sandwich technique in which antiserum to human IgE is adsorbed on a cellulose acetate/nitrate disc which is attached to a plastic StiQTM sampler. The prepared sampler is reacted with serum and antigen is bound specifically. After buffer wash and treatment with goat serum to block non-specific binding, the samplers are reacted with monospecific fluorescein conjugated antisera to human IgE. Two buffer washes remove unbound material and the fluorescence which is directly proportional to the IgE concentration is measured in a FIAX fluorometer. Assay standards range from 2 to 400 IU/ml. The method gives within run coefficients of variation (CV) from 3.3 to 8.4% and between run CV from 6.2 to 16.1% being less precise at low analyte concentrations. IgE concentrations of a group of 74 sera determined by FIA using StiQsTM prepared with antisera from 2 sources correlated well with results found by radioimmunoassay.
本文描述了一种用于测量免疫球蛋白E（IgE）的荧光免疫分析（FIA）方法。该方法采用夹心技术，即将抗人IgE抗血清吸附在附着于塑料StiQTM采样器的醋酸纤维素/硝酸纤维素圆盘上。将制备好的采样器与血清反应，使抗原特异性结合。经过缓冲液洗涤并用山羊血清处理以阻断非特异性结合后，采样器再与单特异性荧光素标记的抗人IgE抗血清反应。两次缓冲液洗涤去除未结合的物质，然后在FIAX荧光计中测量与IgE浓度成正比的荧光。分析标准品的范围为2至400 IU/ml。该方法批内变异系数（CV）为3.3%至8.4%，批间CV为6.2%至16.1%，在低分析物浓度时精密度较低。使用来自2个来源抗血清制备的StiQsTM通过FIA测定的一组74份血清的IgE浓度与放射免疫分析结果相关性良好。
