Ryall R G, Gjerde E M, Gerace R L, Ranieri E
Department of Chemical Pathology, Adelaide Children's Hospital, South Australia.
Clin Chem. 1993 Feb;39(2):224-8.
A coated microtiter-well, enzyme-linked immunometric assay for quantifying immunoreactive trypsinogen in dried blood spots was modified to use time-resolved fluorescence of europium in place of end-point enzymatic color development as the quantification step. The streptavidin-horseradish peroxidase and color development solutions supplied as packaged reagents were replaced by europium-labeled avidin, and the signal was developed with commercially available enhancement solution and read by time-resolved fluorescence. The change of label from enzyme to europium increased the dynamic range of the assay by about 5-fold, reduced the detection limit 10-fold, and halved the intra- and interassay imprecision. The improved analytical precision and stability of the modified assay resulted in a more precise description of the population distribution of immunoreactive trypsinogen values in newborns, showing less variance in the upper centiles. This effect is of paramount importance when using this assay for neonatal screening for cystic fibrosis.
一种用于定量干血斑中免疫反应性胰蛋白酶原的包被微量滴定板酶联免疫测定法进行了改进，用铕的时间分辨荧光取代终点酶显色作为定量步骤。作为包装试剂提供的链霉亲和素 - 辣根过氧化物酶和显色溶液被铕标记的抗生物素蛋白取代，信号用市售增强溶液显影，并通过时间分辨荧光读取。标记从酶变为铕使测定的动态范围增加了约5倍，检测限降低了10倍，批内和批间不精密度减半。改进后的测定法提高的分析精密度和稳定性，使得对新生儿免疫反应性胰蛋白酶原值的总体分布描述更加精确，在上百分位数中显示出较小的差异。当使用该测定法进行囊性纤维化新生儿筛查时，这种效果至关重要。
