Programa de Pós-Graduação em Inovação Terapêutica, Universidade Federal de Pernambuco, 50670-901 Recife, PE, Brazil; Laboratório de Biodispositivos Nanoestruturados, Departamento de Bioquímica, Universidade Federal de Pernambuco, 50670-901 Recife, PE, Brazil.
MEMS, Microfluidics and Nanoelectronics Laboratory, Department of Electrical and Electronics Engineering, Birla Institute of Technology and Science, Hyderabad, 500078, India.
Bioelectrochemistry. 2024 Dec;160:108795. doi: 10.1016/j.bioelechem.2024.108795. Epub 2024 Aug 13.
E6 and E7 oncogenes are pivotal in the carcinogenic transformation in HPV infections and efficient diagnostic methods can ensure the detection and differentiation of HPV genotype. This study describes the development and validation of an electrochemical, label-free genosensor coupled with a microfluidic system for detecting the E6 and E7 oncogenes in cervical scraping samples. The nanostructuring employed was based on a cysteine and graphene quantum dots layer that provides functional groups, surface area, and interesting electrochemical properties. Biorecognition tests with cervical scraping samples showed differentiation in the voltammetric response. Low-risk HPV exhibited a lower biorecognition response, reflected in ΔI% values of 82.33 % ± 0.29 for HPV06 and 80.65 % ± 0.68 for HPV11 at a dilution of 1:100. Meanwhile, high-risk, HPV16 and HPV18, demonstrated ΔI% values of 96.65 % ± 1.27 and 93 % ± 0.026, respectively, at the same dilution. Therefore, the biorecognition intensity followed the order: HPV16 >HPV18 >HPV06 >HPV11. The limit of detection and the limit of quantification of E6E7 microfluidic LOC-Genosensor was 26 fM, and 79.6 fM. Consequently, the E6E7 biosensor is a valuable alternative for clinical HPV diagnosis, capable of detecting the potential for oncogenic progression even in the early stages of infection.
E6 和 E7 癌基因在 HPV 感染的致癌转化中起着关键作用,有效的诊断方法可以确保 HPV 基因型的检测和区分。本研究描述了一种电化学、无标记的基因传感器与微流控系统的开发和验证,用于检测宫颈刮片样本中的 E6 和 E7 癌基因。所采用的纳米结构基于半胱氨酸和石墨烯量子点层,提供了功能基团、表面积和有趣的电化学性质。用宫颈刮片样本进行的生物识别测试显示出了在伏安响应方面的差异。低危型 HPV 表现出较低的生物识别反应,在 1:100 的稀释度下,HPV06 的 ΔI%值为 82.33%±0.29,HPV11 的 ΔI%值为 80.65%±0.68。相比之下,高危型 HPV16 和 HPV18 的 ΔI%值分别为 96.65%±1.27 和 93%±0.026。因此,生物识别强度的顺序为:HPV16>HPV18>HPV06>HPV11。E6E7 微流控 LOC-Genosensor 的检测限和定量限分别为 26 fM 和 79.6 fM。因此,E6E7 生物传感器是一种有价值的临床 HPV 诊断替代方法,即使在感染的早期阶段,也能够检测到致癌进展的潜力。