LAMP-assisted electrochemical DNA biosensor for extraction-free detection of HPV-16 from cervical cancer cell lysates
MICROCHEMICAL JOURNAL, cilt.229, 2026 (SCI-Expanded, Scopus)
- Yayın Türü: Makale / Tam Makale
- Cilt numarası: 229
- Basım Tarihi: 2026
- Doi Numarası: 10.1016/j.microc.2026.119439
- Dergi Adı: MICROCHEMICAL JOURNAL
- Derginin Tarandığı İndeksler: Science Citation Index Expanded (SCI-EXPANDED), Scopus, BIOSIS, Chemical Abstracts Core, Chimica, Index Islamicus, Academic Search Ultimate (EBSCO), Engineering Source (EBSCO)
- İstanbul Üniversitesi Adresli: Evet
Özet
In this work, a LAMP-integrated electrochemical DNA biosensor is presented for ultrasensitive detection of high-risk human papillomavirus type 16 (HPV-16) directly from crude cervical cancer cell lysates. A conductive polypyrrole (PPy) nanofilm was electropolymerized onto a pencil graphite electrode (PGE) to enhance interfacial charge transfer and provide a stable immobilization platform. A biotinylated capture probe targeting the HPV-16 E7 oncogene was immobilized via an avidin-biotin strategy to promote oriented binding and efficient hybridization. Signal transduction was achieved using methylene blue (MB) as a redox-active indicator through a signal-off mechanism monitored by differential pulse voltammetry, where the electrochemical MB signal decreased with increasing target concentration, demonstrating the characteristic signal-off response of the proposed biosensor. All fabrication steps were systematically characterized, confirming the successful and stepwise construction of the biosensor interface. Under optimized conditions, the biosensor demonstrated a linear response toward a synthetic HPV-16 target oligonucleotide over the concentration range of 10_ 1-10_ 7 mu M, achieving an ultralow detection limit of 0.852 fM. High selectivity was confirmed against non-complementary sequences and HPV-18-positive HeLa cell lysates. Integration with loop-mediated isothermal amplification (LAMP; 65 degrees C, 60 min) enabled direct detection of HPV-16 from thermally lysed (95 degrees C, 10 min) SiHa and CaSki cells without DNA extraction or thermal denaturation. The platform demonstrated excellent reproducibility and retained 92.6% of its initial signal after 75 days of storage. By combining isothermal amplification with a nanostructured electrochemical interface and denaturation-free hybridization, this work presents a simplified yet highly sensitive strategy for HPV-16 detection, with a sample-to-result time of approximately 125 min and strong potential for point-of-care molecular diagnostics.