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一种新型定量双抗原夹心 ELISA 法检测白念珠菌烯醇酶 1 总抗体。

A novel quantitative double antigen sandwich ELISA for detecting total antibodies against Candida albicans enolase 1.

机构信息

Department of Nephrology, Bethune International Peace Hospital, No.398 West Zhongshan Road, Shijiazhuang, Hebei, 050082, P.R. China.

Blood Transfusion, Southern Theater General Hospital, No.111 Liuhua Road, Guangzhou, 510180, P.R. China.

出版信息

Eur J Clin Microbiol Infect Dis. 2024 Sep;43(9):1815-1823. doi: 10.1007/s10096-024-04899-4. Epub 2024 Jul 16.

Abstract

PURPOSE

This study aimed to develop a double antigen sandwich ELISA (DAgS-ELISA) method for more efficient, accurate, and quantitative detection of total antibodies against Candida albicans enolase1 (CaEno1) for diagnosing invasive candidiasis (IC).

METHODS

DAgS-ELISA was developed using recombinant CaEno1 and a monoclonal antibody as the standard. Performance evaluation included limit of detection, accuracy, and repeatability. Dynamic changes in antibody levels against CaEno1 in serum from systemic candidiasis mice were analyzed using DAgS-ELISA. Patient serum samples from IC, Candida colonization, bacterial infections, and healthy controls were analyzed with DAgS-ELISA and indirect ELISA.

RESULTS

DAgS-ELISA outperformed indirect ELISA in terms of linear range and test background. In systemic candidiasis mice, a distinctive 'double-peak' pattern in dynamic antibody levels was observed. Additionally, there was a high level of consistency in the positive rates of CaEno1 antibodies detected by both DAgS-ELISA and indirect ELISA. While the positivity rates differed among patient groups, no significant variations in antibody levels were detected among the various positive patient groups.

CONCLUSIONS

DAgS-ELISA offers a reliable novel approach for IC diagnosis, enabling rapid, accurate, and quantitative detection of CaEno1 antibodies. Further validation and optimization are needed for its clinical application and effectiveness.

摘要

目的

本研究旨在开发一种双抗原夹心 ELISA(DAgS-ELISA)方法,用于更有效地、准确地、定量检测针对白色念珠菌烯醇酶 1(CaEno1)的总抗体,以诊断侵袭性念珠菌病(IC)。

方法

使用重组 CaEno1 和单克隆抗体作为标准来开发 DAgS-ELISA。性能评估包括检测限、准确性和重复性。使用 DAgS-ELISA 分析系统性念珠菌病小鼠血清中针对 CaEno1 的抗体水平的动态变化。使用 DAgS-ELISA 和间接 ELISA 分析 IC、念珠菌定植、细菌感染和健康对照患者的血清样本。

结果

DAgS-ELISA 在线性范围和测试背景方面优于间接 ELISA。在系统性念珠菌病小鼠中,观察到动态抗体水平的独特“双峰”模式。此外,DAgS-ELISA 和间接 ELISA 检测的 CaEno1 抗体阳性率高度一致。虽然患者组的阳性率不同,但不同阳性患者组的抗体水平没有差异。

结论

DAgS-ELISA 为 IC 诊断提供了一种可靠的新方法,能够快速、准确、定量地检测 CaEno1 抗体。需要进一步验证和优化其临床应用和效果。

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