Metal-polymer hybrid nanomaterial for impedimetric detection of human papillomavirus in cervical specimens

J Pharm Biomed Anal. 2020 Jun 5:185:113249. doi: 10.1016/j.jpba.2020.113249. Epub 2020 Mar 9.

Abstract

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-1 to 100 pg μL-1 range. Besides, a limit of detection (LOD) of 2.74 pg μL-1 and 7.43 pg μL-1 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.

Keywords: Biosensor; Electrochemical impedance spectroscopy; Gold nanoparticle; Human papillomavirus; Polyaniline.

Publication types

  • Comparative Study
  • Observational Study

MeSH terms

  • Alphapapillomavirus / genetics
  • Alphapapillomavirus / isolation & purification*
  • Biosensing Techniques / methods*
  • Cervix Uteri / pathology
  • Cervix Uteri / virology*
  • DNA, Viral / isolation & purification
  • Diagnosis, Differential
  • Feasibility Studies
  • Female
  • Genotyping Techniques / methods
  • Gold / chemistry
  • Humans
  • Limit of Detection
  • Metal Nanoparticles / chemistry*
  • Metal Nanoparticles / ultrastructure
  • Microelectrodes
  • Microscopy, Atomic Force
  • Molecular Probes / chemistry
  • Papillomavirus Infections / diagnosis*
  • Papillomavirus Infections / pathology
  • Papillomavirus Infections / virology
  • Prognosis
  • Uterine Cervical Neoplasms / prevention & control
  • Uterine Cervical Neoplasms / virology

Substances

  • DNA, Viral
  • Molecular Probes
  • Gold