Department of Veterinary Science, University of Pisa, 56122, Via Livornese, (PI), Italy.
Department of Chemistry "Ugo Schiff", University of Florence, 50019, Sesto Fiorentino, (FI), Italy.
Talanta. 2021 Aug 1;230:122347. doi: 10.1016/j.talanta.2021.122347. Epub 2021 Mar 23.
Procalcitonin (PCT) has emerged as a promising biomarker for the rapid identification of sepsis both in human and veterinary medicine. Nevertheless, the only analytical method currently available for the detection of PCT in veterinary species, is represented by immunoassays, useful only for research purposes. In this work, we report the development of two biosensors which utilize molecularly imprinted polymers (MIPs) for the detection of canine and equine PCT. Dopamine (DA) and norepinephrine (NE) were used as monomers for the synthesis of the MIP films on surface plasmon resonance (SPR) gold chips and the imprinting efficiency of canine and equine PCT in terms of binding affinity toward the analyte, selectivity, and sensitivity were compared. After optimization in buffer conditions, PCTs calibration was successfully achieved also in animal plasma, with good specificity and reproducibility. More effective protein binding and imprinting was obtained with polynorepinephrine (PNE) for both PCTs, and the SPR biosensors were able to detect the biomarkers in plasma with a LOD of 15 ng mL and 30 ng mL respectively for equine and canine PCT.
降钙素原（PCT）已成为一种很有前途的生物标志物，可用于快速识别人类和兽医医学中的败血症。然而，目前兽医物种中检测 PCT 的唯一分析方法是免疫测定法，仅可用于研究目的。在这项工作中，我们报告了两种生物传感器的开发，该传感器利用分子印迹聚合物（MIP）用于检测犬和马的 PCT。多巴胺（DA）和去甲肾上腺素（NE）被用作合成表面等离子体共振（SPR）金芯片上的 MIP 膜的单体，并且比较了犬和马 PCT 的结合亲和力、选择性和灵敏度方面的印迹效率。在缓冲条件下进行优化后，还成功地在动物血浆中实现了 PCT 的校准，具有良好的特异性和重现性。对于两种 PCT 来说，多去甲肾上腺素（PNE）都能获得更好的蛋白结合和印迹效果，SPR 生物传感器能够检测到血浆中的生物标志物，马和犬的 PCT 的检测限分别为 15ng/mL 和 30ng/mL。
