Human papillomavirus type 16 (HPV16) is a significant biomarker for cervical cancer, and its precise detection is vital for diagnosing the disease. The work involved developing an electrochemical sensor by utilizing copper nanoclusters (CuNCs), which integrated triblock polyadenine probe (TPP) and hybridization chain reaction (HCR) for sensitively detecting HPV16 DNA. The TPP was assembled on the electrode owing to the strong affinity between its adenine bases and gold. The absence of HPV16 DNA allowed the capture probe to bind with the hairpin DNA, which triggered HCR and generated a large number of CuNCs, causing a substantial electrochemical impedance signal. However, when there was HPV16 DNA in the reaction system, it could bind specifically to the two flanking probes of TPP, causing the release of the capture probe. Without capture probe, the HCR was unable to be triggered, thus generating a small amount of CuNCs and resulting in a reduced electrochemical impedance signal. With the optimal conditions, the sensor could detect HPV16 DNA within a broad linear range of 1 × 10 −12 M – 1 × 10 −5 M and had a low detection limit of 2.6 × 10 −13 M. A new analytical platform for the sensitive detection of DNA was developed, which exhibited good sensitivity and selectivity.
Human papillomavirus (HPV) is the primary cause of cervical cancer, a major global health concern affecting women worldwide. Early detection of high-risk HPV genotypes is crucial for timely intervention and reducing disease burden. However, current detection methods often lack sensitivity, speed, or cost-effectiveness, especially in resource-limited settings. This work addresses this critical need by developing an ultrasensitive electrochemical biosensing platform based on nitrogen-doped graphene paper electrodes for quantifying high-risk HPV16 DNA sequences. The N-graphene nanohybrid, fabricated via simple solvothermal treatment of biomass-derived graphene oxide, demonstrated significantly enhanced electrical and electrocatalytic properties compared to undoped graphene. DNA detection was achieved through characteristic oxidation signals of guanine bases, which displayed a wide linear dynamic range (0.5–100 μg/mL), low detection limit (75 pg/mL), and excellent reproducibility (relative standard deviation <3.5 %). Detailed spectroscopic and electrochemical analyses revealed the key roles of pyridinic nitrogen defects in improving interfacial charge transfer kinetics and mitigating biofouling issues. This synergistic combination of high electrical conductivity and versatile surface functionality paves the way for equipment-free, point-of-care genosensor devices tailored for quantitative biomarker screening in resource-constrained environments.
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