Glória Juliane Corrêa, Oliveira Daniela S, Gandarilla Ariamna Dip, Barcelay Yonny Romaguera, Mariúba Luis André Morais, Nogueira Paulo Afonso, Brito Walter Ricardo, Moreira Felismina T C
Programa de Pós-graduação em Biotecnologia da Universidade Federal do Amazonas - UFAM, Manaus, AM 69077-000, Brazil.
Instituto Leônidas e Maria Deane (ILMD), Fundação Oswaldo Cruz (FIOCRUZ), Manaus, AM 69057-070, Brazil.
ACS Omega. 2024 Jul 15;9(30):33130-33139. doi: 10.1021/acsomega.4c04543. eCollection 2024 Jul 30.
Malaria is a major public health challenge worldwide and requires accurate and efficient diagnostic methods. Traditional diagnostic approaches based on antigen-antibody interactions are associated with ethical and economic concerns. Molecularly imprinted polymers (MIPs) offer a promising alternative by providing a complementary polymer structure capable of selectively binding target molecules. In this study, we developed a liquid, redox-probe-free, MIP-based electrochemical biosensor to detect the malaria marker histidine-rich protein (HRP2) at the point-of-care (PoC). The imprinting phase consists of the electropolymerization of the monomer methylene blue (MB) in the presence of the target protein HRP2 at the working electrode (WE) of the modified carbon screen printed electrode (C-SPE). Subsequent removal of the protein with proteinase K and oxalic acid yielded the MIP material. The sensor assembly was monitored by cyclic voltammetry (CV), Raman spectroscopy and scanning electron microscopy (SEM). The analytical performance of the biosensor was evaluated by square-wave voltammetry (SWV) using calibration curves in buffer and serum with a detection limit of 0.43 ± 0.026 pg mL. Selectivity studies showed minimal interference, indicating a highly selective assay. Overall, our approach to detect the HRP2 infection marker offers simplicity, cost-effectiveness and reliability. In particular, the absence of a redox solution simplifies detection, as the polymer itself is electroactive and exhibits oxidation and reduction peaks.
疟疾是全球主要的公共卫生挑战，需要准确高效的诊断方法。基于抗原 - 抗体相互作用的传统诊断方法存在伦理和经济方面的问题。分子印迹聚合物（MIPs）通过提供一种能够选择性结合目标分子的互补聚合物结构，提供了一种有前景的替代方案。在本研究中，我们开发了一种基于MIP的液体、无氧化还原探针的电化学生物传感器，用于在护理点（PoC）检测疟疾标志物富含组氨酸蛋白（HRP2）。印迹阶段包括在修饰的碳丝网印刷电极（C - SPE）的工作电极（WE）上，在目标蛋白HRP2存在下，单体亚甲基蓝（MB）的电聚合。随后用蛋白酶K和草酸去除蛋白质，得到MIP材料。通过循环伏安法（CV）、拉曼光谱和扫描电子显微镜（SEM）对传感器组件进行监测。使用缓冲液和血清中的校准曲线，通过方波伏安法（SWV）评估生物传感器的分析性能，检测限为0.43±0.026 pg/mL。选择性研究表明干扰最小，表明该检测方法具有高度选择性。总体而言，我们检测HRP2感染标志物的方法具有简单、成本效益高和可靠性的特点。特别是，由于聚合物本身具有电活性并呈现氧化和还原峰，无氧化还原溶液简化了检测过程。
