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.
J Pharm Biomed Anal. 2020 Jun 5;185:113249. doi: 10.1016/j.jpba.2020.113249. Epub 2020 Mar 9.
The human papillomavirus (HPV) is one of the main sexually transmitted pathogens that infect the anogenital epithelium and mucous membranes. HPV genotypes can be classified as high and low oncogenic risk, with infection by the former resulting in cervical cancer in approximately 100 % of the cases. In this work, we developed an ultrasensitive electrochemical biosensor for the detection and identification of different HPV genotypes. A nanostructured platform based on a matrix of polyaniline (PANI) containing gold nanoparticles (AuNps) was designed for the chemical immobilization of a DNA probe capable of recognizing different HPV types. Cyclic voltammetry (CV), electrochemical impedance spectroscopy (EIS), and atomic force microscopy (AFM) were used to characterize the genosensor. The impedimetric responses indicate that the proposed sensor was able to detect HPV (types 6, 11, 16, 31, 33, 45, and 58) in cervical specimens (cDNA samples). We obtained different profiles of electrochemical responses for the high and low-risk HPV genotypes. By adopting a three-dimensional quantitative analysis of impedance response variables, it was possible to identify the existence of a pattern of association for samples of high oncogenic risk, which may lead to the differential diagnosis of HPV. The biosensor demonstrated an excellent analytical performance for the detection of HPV genotypes with high sensibility and selectivity. The genosensor exhibited a linear range of response in the 1 pg μL to 100 pg μL range. Besides, a limit of detection (LOD) of 2.74 pg μL and 7.43 pg μL was obtained for HPV11 and HPV16, respectively, with regression coefficients of 99.88 % and 99.47 %. Thus, the proposed sensor may serve as a good prognostic indicator for patients infected with papillomavirus.
人乳头瘤病毒（HPV）是一种主要的性传播病原体，可感染肛门生殖器上皮和粘膜。HPV 基因型可分为高和低致癌风险型，前者感染约 100%导致宫颈癌。在这项工作中，我们开发了一种用于检测和识别不同 HPV 基因型的超灵敏电化学生物传感器。设计了一种基于聚苯胺（PANI）基质的纳米结构平台，该基质含有金纳米颗粒（AuNps），用于化学固定能够识别不同 HPV 型的 DNA 探针。循环伏安法（CV）、电化学阻抗谱（EIS）和原子力显微镜（AFM）用于对基因传感器进行表征。阻抗响应表明，所提出的传感器能够检测宫颈标本（cDNA 样本）中的 HPV（类型 6、11、16、31、33、45 和 58）。我们为高低致癌风险 HPV 基因型获得了不同的电化学响应曲线。通过采用阻抗响应变量的三维定量分析，可以识别出高致癌风险样本存在关联模式，这可能导致 HPV 的鉴别诊断。生物传感器对 HPV 基因型的检测具有出色的分析性能，具有高灵敏度和选择性。该基因传感器在 1 pg μL 至 100 pg μL 的范围内呈现出线性响应范围。此外，对于 HPV11 和 HPV16，分别获得了 2.74 pg μL 和 7.43 pg μL 的检测限（LOD），回归系数分别为 99.88%和 99.47%。因此，该传感器可能成为感染 HPV 的患者的良好预后指标。
