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上转换发光高灵敏度免疫分析法检测心脏肌钙蛋白 T 长片段

Highly Sensitive Immunoassay for Long Forms of Cardiac Troponin T Using Upconversion Luminescence.

机构信息

Biotechnology Unit, Department of Life Technologies, University of Turku, Turku, Finland.

Heart Center, Turku University Hospital and University of Turku, Turku, Finland.

出版信息

Clin Chem. 2024 Aug 1;70(8):1037-1045. doi: 10.1093/clinchem/hvae075.

Abstract

BACKGROUND

Long cardiac troponin T (cTnT) has been proposed to be a promising and more specific biomarker of acute myocardial infarction (AMI). As it represents a subfraction of circulating cTnT, detection of very low concentrations is a requirement. The aim of this study was to develop a novel, highly sensitive immunoassay for long cTnT.

METHODS

A two-step sandwich-type immunoassay for long cTnT was developed, utilizing upconverting nanoparticles (UCNPs) as reporters. The limits of detection and quantitation were determined for the assay. Linearity and matrix effects were evaluated. Performance with clinical samples was assessed with samples from patients with non-ST elevation myocardial infarction (NSTEMI, n = 30) and end-stage renal disease (ESRD, n = 37) and compared to a previously developed time-resolved fluorescence (TRF)-based long cTnT assay and a commercial high-sensitivity cTnT assay.

RESULTS

The novel assay reached a 28-fold lower limit of detection (0.40 ng/L) and 14-fold lower limit of quantitation (1.79 ng/L) than the previously developed TRF long cTnT assay. Li-heparin and EDTA plasma, but not serum, were found to be suitable sample matrixes for the assay. In a receiver operating characteristics curve analysis, the troponin ratio (long/total cTnT) determined with the novel assay showed excellent discrimination between NSTEMI and ESRD with an area under the curve of 0.986 (95% CI, 0.967-1.000).

CONCLUSIONS

By utilizing upconversion luminescence technology, we developed a highly sensitive long cTnT assay. This novel assay can be a valuable tool for investigating the full potential of long cTnT as a biomarker for AMI. ClinicalTrials.gov Registration Number: NCT04465591.

摘要

背景

长型心肌肌钙蛋白 T(cTnT)被认为是急性心肌梗死(AMI)有前途且更具特异性的生物标志物。由于它代表循环 cTnT 的亚片段,因此需要检测非常低的浓度。本研究旨在开发一种新型的高灵敏度长型 cTnT 免疫测定法。

方法

开发了一种两步夹心型长型 cTnT 免疫测定法,利用上转换纳米粒子(UCNPs)作为报告器。确定了该测定法的检测限和定量限。评估了线性度和基质效应。使用非 ST 段抬高型心肌梗死(NSTEMI,n=30)和终末期肾病(ESRD,n=37)患者的样本评估了与之前开发的基于时间分辨荧光(TRF)的长型 cTnT 测定法和商业高灵敏度 cTnT 测定法的临床样本的性能。

结果

与之前开发的 TRF 长型 cTnT 测定法相比,新型测定法的检测限降低了 28 倍(0.40ng/L),定量限降低了 14 倍(1.79ng/L)。发现 Li-heparin 和 EDTA 血浆,但不是血清,是该测定法合适的样本基质。在受试者工作特征曲线分析中,新型测定法确定的肌钙蛋白比值(长型/total cTnT)在 NSTEMI 和 ESRD 之间具有出色的区分能力,曲线下面积为 0.986(95%CI,0.967-1.000)。

结论

通过利用上转换发光技术,我们开发了一种高灵敏度的长型 cTnT 测定法。这种新型测定法可以成为研究长型 cTnT 作为 AMI 生物标志物的全部潜力的有价值的工具。临床试验注册编号:NCT04465591。

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