Microfluidic-Based Genosensor To Detect Human Papillomavirus (HPV16) for Head and Neck Cancer

• Published in: ACS applied materials & interfaces Vol. 10; no. 43; pp. 36757 - 36763
• Main Authors: Soares, Andrey Coatrini, Soares, Juliana Coatrini, Rodrigues, Valquiria Cruz, Follmann, Heveline Dal Magro, Arantes, Lidia Maria Rebolho Batista, Carvalho, Ana Carolina, Melendez, Matias Eliseo, Fregnani, José Humberto T. G, Reis, Rui Manuel, Carvalho, André Lopes, Oliveira, Osvaldo N
• Format: Journal Article
• Language: English
• Published: United States American Chemical Society 31.10.2018
• Subjects: Adenine - chemistry; Carcinoma, Squamous Cell - diagnosis; Carcinoma, Squamous Cell - virology; Cell Line, Tumor; Chitosan - chemistry; Chondroitin Sulfates - chemistry; DNA, Single-Stranded - chemistry; Electric Impedance; Head and Neck Neoplasms - diagnosis; Head and Neck Neoplasms - virology; Human papillomavirus 16 - isolation & purification; Humans; Limit of Detection; Microfluidic Analytical Techniques; Nanostructures - chemistry; Papillomavirus Infections - diagnosis; Thymine - chemistry; impedance spectroscopy; genosensors; head and neck cancer; layer-by-layer films; multidimensional projections; HPV
• DOI: 10.1021/acsami.8b14632
• ISSN: 1944-8244

High-risk human papillomavirus (HPV) infection, mainly with HPV16 type, has been increasingly considered as an important etiologic factor in head and neck cancers. Detection of HPV16 is therefore crucial for these types of cancer, but clinical tests are not performed routinely in public health systems owing to the high cost and limitations of the existing tests. In this article, we report on a potentially low-cost genosensor capable of detecting low concentrations of HPV16 in buffer samples and distinguishing, with high accuracy, head and neck cancer cell lines according to their HPV16 status. The genosensor consisted of a microfluidic device that had an active layer of a HPV16 capture DNA probe (cpHPV16) deposited onto a layer-by-layer film of chitosan and chondroitin sulfate. Impedance spectroscopy was the principle of detection utilized, leading to a limit of detection of 10.5 pM for complementary ssDNA HPV16 oligos (ssHPV16). The genosensor was also able to distinguish among HPV16+ and HPV16– cell lines, using the multidimensional projection technique interactive document mapping. Hybridization between the ssHPV16 oligos and cpHPV16 probe was confirmed with polarization-modulated infrared reflection-absorption spectroscopy, where PO2 and amide I and amide II bands from adenine and thymine were monitored. The electrical response could be modeled as resulting from an adsorption process represented in a Freundlich model. Because the fabrication procedures of the microfluidic devices and genosensors and the data collection and analysis can be implemented at low cost, the results presented here amount to a demonstration of possible routine screening for HPV infections.