A quantitative enzyme-linked immunoassay (ELISA) to approximate complement-fixing antibody titers in serum from patients with coccidioidomycosis
A quantitative enzyme-linked immunoassay (ELISA) to approximate complement-fixing antibody titers in serum from patients with coccidioidomycosis
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DOI：
10.1016/j.diagmicrobio.2020.115198
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发表时间：
2021-01-01
影响因子：
2.9
通讯作者：
Galgiani, John N.
中科院分区：
文献类型：
--
作者：
Peng, Tao;Zong, Yue;Galgiani, John N.
球孢子菌病最常通过血清学诊断，补体固定抗体的定量检测被认为对预后有用。由于补体固定抗体检测复杂、劳动强度大且标准化程度差，因此酶联免疫分析 (ELISA) 替代方法将很有吸引力。在本报告中，我们将补体固定抗体结合结构域限制为已知抗原的 200 个氨基酸重组肽。该肽的重叠截短不结合补体固定抗体，表明负责的表位是构象的。此外，通过C端生物素模拟肽标签将抗原肽锚定至ELISA板，而不是让肽随机粘附至塑料板，从而将不同血清中抗体检测的灵敏度提高了1-2个对数。新开发的 ELISA 显示与补体固定抗体滴度具有显着的定量相关性。该 ELISA 显示出作为球孢子抗体新定量测定基础的潜力。 (C) 2020 Elsevier Inc. 保留所有权利。
Coccidioidomycosis is most frequently diagnosed serologically, and the quantitative test for complement-fixing antibodies is considered prognostically useful. Because complement-fixing antibody testing is complex, laborintensive, and poorly standardized, an enzyme-linked immunoassay (ELISA) alternative would be attractive. In this report, we restrict the complement-fixing, antibody-binding domain to a 200-amino-acid recombinant peptide of the known antigen. Over-lapping truncations of this peptide do not bind complement-fixing antibodies, suggesting that the responsible epitope(s) are conformational. Further, anchoring the antigenic peptide to the ELISA plate by means of a C-terminal biotin-mimic peptide tag instead of allowing the peptide to randomly adhere to the plastic plate improves sensitivity of antibody detection by 1-2 logs in different sera. The newly developed ELISA shows a significant quantitative correlation with complement-fixing antibody titers. This ELISA shows potential as the basis for a new quantitative assay for coccidioidal antibodies. (C) 2020 Elsevier Inc. All rights reserved.