Detection of HPV16 in cell lines deriving from cervical and head and neck cancer using a genosensor made with a DNA probe on a layer-by-layer matrix

Author(s)

Soares, Juliana Coatrini ^{[1, 2]} ; Melendez, Matias Eliseo ^{[3, 4, 5]} ; Soares, Andrey Coatrini ^[1] ; Arantes, Lidia Maria Rebolho Batista ^[3] ; Rodrigues, Valquiria da Cruz ^{[6, 2]} ; Carvalho, Andre Lopes ^[3] ; Reis, Rui Manuel ^{[3, 7]} ; Mattoso, Luiz Henrique C. ^[1] ; Oliveira, Jr., Osvaldo N. ^[2]

Affiliation

^[1] Embrapa Instrumentacao, Nanotechnol Natl Lab Agr LNNA, BR-13560970 Sao Carlos, SP - Brazil

^[2] Univ Sao Paulo, Sao Carlos Inst Phys, BR-13566590 Sao Carlos - Brazil

^[3] Barretos Canc Hosp, Mol Oncol Res Ctr, BR-14784400 Barretos - Brazil

^[4] Fac Pequeno Principe, Curitiba, PR - Brazil

^[5] Pele Pequeno Principe Res Inst, Curitiba, PR - Brazil

^[6] Univ Sao Paulo, Sao Carlos Sch Engn, BR-13566590 Sao Carlos - Brazil

^[7] Univ Minho, Sch Med, Life & Hlth Sci Res Inst ICVS, P-4710057 Braga - Portugal

• Document type: Journal article
• Source: MATERIALS CHEMISTRY FRONTIERS; v. 4, n. 11, p. 3258-3266, NOV 1 2020.
• Web of Science Citations: 0

Abstract

Detection of human papillomavirus 16 (HPV16) is essential for the diagnosis of cervix and head and neck cancer, which has prompted the development of biosensors to replace the sophisticated molecular techniques. In this paper, we employ a genosensor made with a HPV16 DNA probe (cpHPV16) immobilized on a layer-by-layer (LbL) film of chitosan and carbon nanotubes to detect a complementary sequence HPV16 oligo (ssHPV16). In addition to employing impedance spectroscopy, which has already been used in the literature, we detect ssHPV16 in buffer samples and in cancer cell line samples with UV-vis. spectroscopy. The limit of detection was 18.5 pmol L-1 in impedance spectroscopy measurements and 10.9 pmol L-1 in UV-vis measurements. Distinction of the various ssHPV16 concentrations and of the cancer cell line samples CasKi, SiHa, UMSCC-47, UM-SCC104 and 93-VU147T with both electrical and optical data could be made with the multidimensional projection technique referred to as interactive document mapping (IDMAP), with which we could confirm the absence of false positives by testing cell lines that did not contain ssHPV16 (JHU12, JHU13 and JHU28). The hybridization between cpHPV16 and ssHPV16 responsible for the biosensing was verified with polarization-modulated infrared reflection absorption spectroscopy (PM-IRRAS), while the concentration dependence could be modeled with a double Freundlich function. The use of optical absorbance measurements to detect ssHPV16 is promising toward a simple colorimetric detection. (AU)

• FAPESP's process: 18/18953-8 - Nanostructured films applied in microfluidic biosensors to mastitis detection
• Grantee: Andrey Coatrini Soares
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• FAPESP's process: 18/22214-6 - Towards a convergence of technologies: from sensing and biosensing to information visualization and machine learning for data analysis in clinical diagnosis
• Grantee: Osvaldo Novais de Oliveira Junior
• Support Opportunities: Research Projects - Thematic Grants