用于检测人血液中地高辛的间接竞争酶联免疫吸附测定法和侧向流动免疫层析试纸条的开发。
Development of Indirect Competitive Enzyme-Linked Immunosorbent Assay and Lateral-Flow Immunochromatographic Strip for the Detection of Digoxin in Human Blood.
作者信息
Ling Fanqian, Liu Liqiang, Kuang Hua, Cui Gang, Xu Chuanlai
机构信息
State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi 214122, People's Republic of China.
International Joint Research Laboratory for Biointerface and Biodetection and School of Food Science and Technology, Jiangnan University, Wuxi 214122, People's Republic of China.
出版信息
ACS Omega. 2020 Jan 13;5(3):1371-1376. doi: 10.1021/acsomega.9b02254. eCollection 2020 Jan 28.
Mouse-mouse hybridoma cell lines producing stable, highly specific monoclonal antibodies with good affinity for the cardiac glycoside digoxin (DIG) were established to construct an indirect enzyme-linked immunosorbent assay and lateral-flow immunochromatographic strip to detect DIG in human blood. The hapten DIG was coupled to bovine serum albumin or chicken ovalbumin by sodium periodate oxidation. The highest sensitivity and specificity antibody had a median inhibitory concentration (IC) of 0.45 ng/mL, a linear range of detection of 0.293-0.7 ng/mL, and low cross-reactivity with several DIG analogues. The cut-off value of the lateral-flow immunochromatographic strip was 5 ng/mL when the strip was tested with human blood. The immunochromatographic lateral flow strip test provides a quick and convenient method for determining DIG in plasma which can be visually observed in only 5 min to promote rational drug use.
建立了能产生对强心苷地高辛（DIG）具有稳定、高特异性且亲和力良好的单克隆抗体的小鼠-小鼠杂交瘤细胞系，以构建间接酶联免疫吸附测定法和侧向流动免疫层析试纸条来检测人血液中的地高辛。半抗原地高辛通过高碘酸钠氧化法与牛血清白蛋白或鸡卵清蛋白偶联。具有最高灵敏度和特异性的抗体的半数抑制浓度（IC）为0.45 ng/mL，检测线性范围为0.293 - 0.7 ng/mL，与几种地高辛类似物的交叉反应性较低。用人血检测试纸条时，侧向流动免疫层析试纸条的临界值为5 ng/mL。免疫层析侧向流动试纸条检测法提供了一种快速便捷的方法来测定血浆中的地高辛，仅需5分钟即可直观观察结果，有助于促进合理用药。
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