Enhanced Paper-Based DNA Adsorption and Point-of-Care Detection of High-Risk Human Papillomaviruses 16 and 18.
5 affiliations
- 1Department of Biotechnology
- 2Indian Institute of Technology Madras
- 3Department of Chemical Engineering
- 4Institute of Obstetrics and Gynaecology (IOG
- 5Madras Medical College
Abstract
Cervical cancer screening in low-resource settings requires molecular diagnostics that integrate rapid DNA extraction with sensitive, low-cost DNA amplification and detection. Here, we present a paper-based DNA detection (PDD) platform that introduces two mechanistic advances for decentralized HPV diagnostics. First, we establish a centrifuge-free DNA isolation strategy based on ethanol-assisted capture of DNA onto cellulose paper to provide high-yield DNA. Second, we introduce a coffee-ring-based fluorescence-sensing mechanism that converts the spatial redistribution of PicoGreen-labeled amplified products into a quantitative readout. We report, for the first time, that acid-pretreated cellulose (APC) paper captures DNA efficiently relative to untreated paper, identifying substrate pH as a critical factor in paper-based DNA extraction. FTIR analysis and zeta potential measurements revealed that APC paper retained DNA efficiently primarily through hydrogen-bonding and electrostatic interactions, respectively. Moreover, the mechanism behind the coffee ring-based amplified DNA detection was attributed to the chromatographic and evaporation effects. DNA is quantified using peripheral fluorescence intensity (PFI), enabling robust discrimination between positive and negative reactions. The integrated platform detected HPV16 and HPV18 with a limit of detection of 10 copies μL⁻¹. Clinical evaluation of 52 cervical samples demonstrated 100% specificity for both HPV16 and HPV18, 100% sensitivity for HPV18, and 85.7% sensitivity for HPV16 compared with an IVD-approved real-time PCR assay. By combining mechanistically guided DNA isolation with physics-based spatial DNA detection, this work advances paper-based molecular diagnostics beyond conventional intensity-based assays and provides a simple, inexpensive, and field-deployable strategy for point-of-care detection of HR HPVs.
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BibTeX
@misc{das2026enhanced,
title = {{Enhanced Paper-Based DNA Adsorption and Point-of-Care Detection of High-Risk Human Papillomaviruses 16 and 18.}},
author = {Neha Rani Das and Mohamed Asif Shajahan and Sivie Kurian and Veeramani Velayutham and Saranya Gopalakrishnan and Shivam Kumar Jha and Sumathi Chandran and Kasthuri Vasu and Vijay Sathish Kumar Irulappan and Vidhya Amirthalingam and Revathy Mahendran and Subramaniam Pushpavanam and Richa Karmakar},
howpublished = {ChemRxiv},
year = {2026},
month = oct,
doi = {10.26434/chemrxiv.15010961/v1},
url = {https://chemrxiv.org/doi/full/10.26434/chemrxiv.15010961/v1},
note = {CC BY},
}
RIS
TY - UNPB
TI - Enhanced Paper-Based DNA Adsorption and Point-of-Care Detection of High-Risk Human Papillomaviruses 16 and 18.
AU - Neha Rani Das
AU - Mohamed Asif Shajahan
AU - Sivie Kurian
AU - Veeramani Velayutham
AU - Saranya Gopalakrishnan
AU - Shivam Kumar Jha
AU - Sumathi Chandran
AU - Kasthuri Vasu
AU - Vijay Sathish Kumar Irulappan
AU - Vidhya Amirthalingam
AU - Revathy Mahendran
AU - Subramaniam Pushpavanam
AU - Richa Karmakar
PY - 2026
DA - 2026/10/23
PB - ChemRxiv
DO - 10.26434/chemrxiv.15010961/v1
UR - https://chemrxiv.org/doi/full/10.26434/chemrxiv.15010961/v1
AB - Cervical cancer screening in low-resource settings requires molecular diagnostics that integrate rapid DNA extraction with sensitive, low-cost DNA amplification and detection. Here, we present a paper-based DNA detection (PDD) platform that introduces two mechanistic advances for decentralized HPV diagnostics. First, we establish a centrifuge-free DNA isolation strategy based on ethanol-assisted capture of DNA onto cellulose paper to provide high-yield DNA. Second, we introduce a coffee-ring-based fluorescence-sensing mechanism that converts the spatial redistribution of PicoGreen-labeled amplified products into a quantitative readout. We report, for the first time, that acid-pretreated cellulose (APC) paper captures DNA efficiently relative to untreated paper, identifying substrate pH as a critical factor in paper-based DNA extraction. FTIR analysis and zeta potential measurements revealed that APC paper retained DNA efficiently primarily through hydrogen-bonding and electrostatic interactions, respectively. Moreover, the mechanism behind the coffee ring-based amplified DNA detection was attributed to the chromatographic and evaporation effects. DNA is quantified using peripheral fluorescence intensity (PFI), enabling robust discrimination between positive and negative reactions. The integrated platform detected HPV16 and HPV18 with a limit of detection of 10 copies μL⁻¹. Clinical evaluation of 52 cervical samples demonstrated 100% specificity for both HPV16 and HPV18, 100% sensitivity for HPV18, and 85.7% sensitivity for HPV16 compared with an IVD-approved real-time PCR assay. By combining mechanistically guided DNA isolation with physics-based spatial DNA detection, this work advances paper-based molecular diagnostics beyond conventional intensity-based assays and provides a simple, inexpensive, and field-deployable strategy for point-of-care detection of HR HPVs.
ER -