title: Development of combined DNA-based piezoelectric biosensors for the simultaneous detection and genotyping of high risk Human Papilloma Virus strains.
abstract: BACKGROUND
  Human Papilloma Virus (HPV) is a DNA virus belonging to the Papovavirus family. Genital HPV types have been subdivided into medium-low risk, and high-risk (HPV 16 and 18), frequently associated with cervical cancer. Three DNA-based piezoelectric biosensors were here developed for a quick detection and genotyping of HPV.
  
  
  METHODS
  We developed a method for the detection and genotyping of HPV in human cervical scraping samples based on coupling DNA piezoelectric sensors with Polymerase Chain Reaction (PCR). The novelty of this work was the design and immobilisation of a degenerate probe (chosen in a conserved region of the viral genome) for the simultaneous detection of 16 virus strains and of two specific probes (chosen in a less-conserved region of the viral genome) for genotyping.
  
  
  RESULTS
  The three biosensors were optimised with synthetic oligonucleotides with good reproducibility (HPVdeg CV% (av) 9%, HPV16 CV%(av) 9%; HPV18 CV%(av) 11%) with a detection limit of 50 nM. Cervical scraping samples after PCR amplification (in 40-200 nM range), were tested without the need of label with high selectivity and reproducibility. The results were in agreement with a reference method used in routinary analysis.
  
  
  CONCLUSIONS
  Piezoelectric biosensors have proven to be suitable for detection and genotyping of HPV.
authors.affiliation: Department of Chemistry, University of Florence, Via della Lastruccia 3, 50019, Sesto Fiorentino, Italy.
authors.forename: Daniela
authors.initials: D
authors.lastname: Dell'Atti
authors.forename: Michele
authors.initials: M
authors.lastname: Zavaglia
authors.forename: Sara
authors.initials: S
authors.lastname: Tombelli
authors.forename: Gloria
authors.initials: G
authors.lastname: Bertacca
authors.forename: Andrea O
authors.initials: AO
authors.lastname: Cavazzana
authors.forename: Generoso
authors.initials: G
authors.lastname: Bevilacqua
authors.forename: Maria
authors.initials: M
authors.lastname: Minunni
authors.forename: Marco
authors.initials: M
authors.lastname: Mascini
mesh_terms: D015374:Biosensing Techniques / Q000295:instrumentation / Q000379:methods* / Q000592:standards*; D004247:DNA; D015344:DNA Probes, HPV; D004279:DNA, Viral; D005260:Female; D005838:Genotype; D006801:Humans; D009841:Oligonucleotides; D027383:Papillomaviridae / Q000235:genetics / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
chemical_list: D015344:DNA Probes, HPV; D004279:DNA, Viral; D009841:Oligonucleotides; D004247:DNA