Supplementary Material for: A Simple and Rapid Light-Initiated Chemiluminescence Assay for Quantitation of <b><i>Artemisia</i></b>-Specific Immunoglobulin E

Background: Light-initiated chemiluminescence assay (LICA) is a homogeneous assay that has been successfully used for the quantitation of food allergen-specific immunoglobulin E (sIgE), but not inhaled allergen-sIgE. Simultaneously, current assays used to detect allergen-sIgE are serum consuming and/or time consuming. Hence, we established a method for the quantitation of Artemisia-sIgE based on LICA and verified its performance according to the clinical guideline documents, laying a foundation for the quantitation of inhaled and food allergen-sIgE in parallel on LICA. Methods: The assay was established after optimizing the first incubation time and the dilutions of Artemisia-coated chemibeads, biotinylated goat anti-human IgE, and serum. In order to quantitate Artemisia-sIgE, the calibration curve was established with a high positive serum of known concentration. The assay performance was confirmed per the clinical guideline documents. In addition, the correlation between the results of LICA and capture enzyme-linked immunosorbent assay was evaluated. Results: The developed LICA’s coefficients of variation of repeatability and intermediate precision were 3.20%, 2.14%, and 3.85% and 4.30%, 4.00%, and 4.40%, respectively. The limit of detection was 0.10 kUA/L, and the limit of quantitation was 0.11 kUA/L. The range of linearity was from 0.27 kUA/L to 97.53 kUA/L (r = 0.9968). The correlation coefficient (r) for the correlation analysis between results of LICA and capture ELISA was 0.9087. This assay was successfully applied in 64 human serum samples, showing good sensitivity (82.20%) and specificity (100%). Conclusion: An Artemisia-sIgE quantitation assay based on LICA was successfully established. Its performance satisfied the clinical requirements and could be widely used in clinical laboratories.背景：光激发化学发光分析法（Light-initiated chemiluminescence assay，LICA）是一种均相分析法，已成功应用于食物过敏原特异性免疫球蛋白E（specific immunoglobulin E，sIgE）的定量检测，但尚未用于吸入性过敏原sIgE的定量。当前用于检测过敏原sIgE的方法均存在血清消耗量大且/或耗时较长的缺陷。为此，本研究基于LICA建立了蒿属过敏原sIgE定量检测方法，并依据临床指南文件对其性能进行验证，为在LICA平台上同步定量检测吸入性与食物过敏原sIgE奠定了研究基础。方法：通过优化首次孵育时间，以及蒿属抗原包被化学微球、生物素标记山羊抗人IgE与血清的稀释比例，构建该检测体系。以已知浓度的高阳性血清绘制校准曲线，实现蒿属过敏原sIgE的定量检测。依据临床指南文件对该方法的分析性能进行确认，并评估了LICA与捕获酶联免疫吸附试验（capture enzyme-linked immunosorbent assay）检测结果的相关性。结果：所建立的LICA方法的重复性与中间精密度的变异系数分别为3.20%、2.14%、3.85%与4.30%、4.00%、4.40%。其检测限为0.10 kUA/L，定量限为0.11 kUA/L，线性范围为0.27 kUA/L至97.53 kUA/L（相关系数r=0.9968）。LICA与捕获酶联免疫吸附试验检测结果的相关系数r为0.9087。该方法成功应用于64份人血清样本，展现出良好的检测性能：灵敏度为82.20%，特异性为100%。结论：本研究成功建立了基于LICA的蒿属过敏原sIgE定量检测方法，其分析性能符合临床要求，可在临床实验室中广泛推广应用。