Development of combined DNA-based piezoelectric biosensors for the simultaneous detection and genotyping of high risk Human Papilloma Virus strains.
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.
Dell'Atti
Daniela
D
Department of Chemistry, University of Florence, Via della Lastruccia 3, 50019, Sesto Fiorentino, Italy.
Zavaglia
Michele
M
Tombelli
Sara
S
Bertacca
Gloria
G
Cavazzana
Andrea O
AO
Bevilacqua
Generoso
G
Minunni
Maria
M
Mascini
Marco
M
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
D015344:DNA Probes, HPV; D004279:DNA, Viral; D009841:Oligonucleotides; D004247:DNA