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.
Laboratório de Biodispositivos Nanoestruturados, Departamento de Bioquímica, Universidade Federal de Pernambuco, 50670-901, Recife, PE, Brazil.
Talanta. 2021 May 1;226:122118. doi: 10.1016/j.talanta.2021.122118. Epub 2021 Jan 21.
Considering the low sensitivity of cytological exams and high costs of the molecular methods, the development of diagnostic tests for effective diagnosis of HPV infections is a priority. In this work, biosensor composed of polypyrrole (PPy) films and gold nanoparticles (AuNPs) was obtained for specific detection of HPV genotypes. The biosensor was developed by using flexible electrodes based on polyethylene terephthalate (PET) strips coated with indium tin oxide (ITO). Polymeric films and AuNPs were obtained by electrosynthesis. Oligonucleotides sequences modified with functional amino groups were designed to recognize HPV gene families strictly. The modified oligonucleotides were chemically immobilized on the nanostructured platform. Cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS) were used for the analysis of the electrode modification and monitoring of molecular hybridization. Electrochemical changes were observed after exposure of the biosensors to plasmid samples and cervical specimens. The biosensor based on the BSH16 probe showed a linear concentration range for target HPV16 gene detection of 100 pg μL to 1 fg μL. A limit of detection (LOD) of 0.89 pg μL and limit of quantification (LOQ) of 2.70 pg μL were obtained, with a regression coefficient of 0.98. Screening tests on cervical specimens were performed to evaluate the sensibility and specificity for HPV and its viral family. The expression of a biomarker for tumorigenesis (p53 gene) was also monitored. In this work, a flexible system has been successfully developed for label-free detection of HPV families and p53 gene monitoring with high specificity, selectivity, and sensitivity.
鉴于细胞学检查的敏感性低和分子方法的成本高，开发用于有效诊断 HPV 感染的诊断测试是当务之急。在这项工作中，制备了由聚吡咯（PPy）薄膜和金纳米粒子（AuNPs）组成的生物传感器，用于特异性检测 HPV 基因型。该生物传感器是通过使用基于聚对苯二甲酸乙二醇酯（PET）条带的柔性电极并涂有氧化铟锡（ITO）来开发的。通过电合成获得聚合物薄膜和 AuNPs。设计了带有功能氨基的寡核苷酸序列来严格识别 HPV 基因家族。将修饰的寡核苷酸化学固定在纳米结构平台上。循环伏安法（CV）和电化学阻抗谱（EIS）用于分析电极修饰和监测分子杂交。在将生物传感器暴露于质粒样品和宫颈标本后观察到电化学变化。基于 BSH16 探针的生物传感器显示出针对 HPV16 基因检测的线性浓度范围为 100 pg μL 至 1 fg μL。获得了 0.89 pg μL 的检测限（LOD）和 2.70 pg μL 的定量限（LOQ），回归系数为 0.98。对宫颈标本进行了筛选测试，以评估 HPV 及其病毒家族的敏感性和特异性。还监测了肿瘤发生的生物标志物（p53 基因）的表达。在这项工作中，成功开发了一种用于 HPV 家族和 p53 基因监测的无标记检测的柔性系统，具有高特异性、选择性和灵敏度。
