title: Electrochemical chip-based genomagnetic assay for detection of high-risk human papillomavirus DNA.
abstract: Cervical cancer, being the fourth leading cause of cancer death in women worldwide, predominantly originates from a persistent infection with a high-risk human papillomavirus (HPV). Detection of DNA sequences from these high-risk strains, mostly HPV-16 and HPV-18, represents promising strategy for early screening, which would help to identify women with higher risk of cervical cancer. In developing countries, inadequate screening options lead to disproportionately high mortality rates, making a fast and inexpensive detection schemes highly important. Electrochemical sensors and assays offer an alternative to current methods of detection. We developed an electrochemical-chip based assay, in which target HPV DNA is captured via magnetic bead-modified DNA probes, followed by an antidigoxigenin-peroxidase detection system at screen-printed carbon electrode chips, enabling parallel measurements of eight samples simultaneously. We show sensitive detection in attomoles of HPV DNA, selective discrimination between HPV-16 and HPV-18 and good reproducibility. Most importantly, we show application of the assay into both cancer cell lines and cervical smears from patients. The electrochemical results correlated well with standard methods, making this assay potentially applicable in clinical practice.
authors.affiliation: Regional Centre for Applied Molecular Oncology (RECAMO), Masaryk Memorial Cancer Institute, Zluty kopec 7, 656 53 Brno, Czech Republic. Electronic address: martin.bartosik@mou.cz.
authors.forename: Martin
authors.initials: M
authors.lastname: Bartosik
authors.affiliation: Regional Centre for Applied Molecular Oncology (RECAMO), Masaryk Memorial Cancer Institute, Zluty kopec 7, 656 53 Brno, Czech Republic.
authors.forename: Helena
authors.initials: H
authors.lastname: Durikova
authors.affiliation: Regional Centre for Applied Molecular Oncology (RECAMO), Masaryk Memorial Cancer Institute, Zluty kopec 7, 656 53 Brno, Czech Republic.
authors.forename: Borivoj
authors.initials: B
authors.lastname: Vojtesek
authors.affiliation: Brno University Hospital and Masaryk University Medical School, Department of Obstetrics and Gynecology, Obilni trh 11, 602 00 Brno, Czech Republic.
authors.forename: Milan
authors.initials: M
authors.lastname: Anton
authors.affiliation: Brno University Hospital and Masaryk University Medical School, Department of Pathology, Jihlavska 20, 625 00 Brno, Czech Republic.
authors.forename: Eva
authors.initials: E
authors.lastname: Jandakova
authors.affiliation: Regional Centre for Applied Molecular Oncology (RECAMO), Masaryk Memorial Cancer Institute, Zluty kopec 7, 656 53 Brno, Czech Republic.
authors.forename: Roman
authors.initials: R
authors.lastname: Hrstka
mesh_terms: D015374:Biosensing Techniques / Q000379:methods; D045744:Cell Line, Tumor; D002584:Cervix Uteri / Q000821:virology; D004279:DNA, Viral / Q000235:genetics*; D055664:Electrochemical Techniques / Q000379:methods*; D005260:Female; D052162:Human papillomavirus 16 / Q000235:genetics / Q000302:isolation & purification; D052161:Human papillomavirus 18 / Q000235:genetics / Q000302:isolation & purification; D006801:Humans; D027383:Papillomaviridae / Q000235:genetics* / Q000302:isolation & purification; D030361:Papillomavirus Infections / Q000175:diagnosis* / Q000821:virology; D015203:Reproducibility of Results; D002583:Uterine Cervical Neoplasms / Q000175:diagnosis* / Q000821:virology
chemical_list: D004279:DNA, Viral
keywords: Cervical cancer; Electrochemical detection; Electrode chip; HPV; Magnetic beads