Dipartimento di Chimica "Ugo Schiff", Via della Lastruccia 3-13, 50019, Sesto Fiorentino, FI, Italy.
Anal Bioanal Chem. 2019 Nov;411(29):7709-7716. doi: 10.1007/s00216-019-02014-7. Epub 2019 Jul 13.
Early diagnosis of acute myocardial infarction (AMI) is of outmost importance to reduce the mortality rate, and cardiac troponins are considered the gold standard biomarkers of myocardial necrosis. In this scenario, the characterization of two troponin T (TnT)-binding aptamers as viable alternative to antibodies employed on clinical immunoassays is here reported for the first time. Their recognition ability was first investigated through surface plasmon resonance (SPR). Subsequently, an enzyme-linked oligonucleotide assay (ELONA) was developed on common 96-well polystyrene plates, both by direct and sandwich detection strategies for comparison. In both cases, the assay exhibits a detection ability of TnT in the range of low nanomolar but a great advantage on serum interference was obtained by using both aptamers in a sandwich format, with excellent reproducibility and recovery values. Despite the sensitivity needing to be enhanced to the low picomolar range, these results are encouraging for the development of new, low-cost, and rapid antibody-free colorimetric assays for AMI studies based on aptamer-Troponin T recognition.
早期诊断急性心肌梗死（AMI）对于降低死亡率至关重要，心肌肌钙蛋白被认为是心肌坏死的金标准生物标志物。在这种情况下，我们首次报道了两种肌钙蛋白 T（TnT）结合适体作为临床免疫分析中使用的抗体的可行替代物的特征。首先通过表面等离子体共振（SPR）研究了它们的识别能力。随后，通过直接和夹心检测策略在常见的 96 孔聚苯乙烯板上开发了酶联寡核苷酸测定法（ELONA）进行比较。在这两种情况下，该测定法在纳摩尔级范围内均具有检测 TnT 的能力，但通过使用两种适体进行夹心检测，在很大程度上克服了血清干扰，得到了极好的重现性和回收率值。尽管需要将灵敏度提高到皮摩尔级范围，但这些结果令人鼓舞，可用于开发基于适体-TnT 识别的新的、低成本、快速的无抗体比色测定法，用于 AMI 研究。
