Human Papilloma Virus DNA Detection in Clinical Samples Using Fluorogenic Probes Based on Oligonucleotide Gated Nanoporous Anodic Alumina Films.
Por:
Hernández-Montoto A, Aranda MN, Caballos I, López-Palacios A, Tormo-Mas MÁ, Pemán J, Rodríguez MP, Picornell C, Aznar E and Martínez-Máñez R
Publicada:
1 sep 2023
Ahead of Print:
1 jun 2023
Resumen:
In this work, fluorogenic probes based on oligonucleotide capped nanoporous anodic alumina films are developed for specific and sensitive detection of human papilloma virus (HPV) DNA. The probe consists of anodic alumina nanoporous films loaded with the fluorophore rhodamine B (RhB) and capped with oligonucleotides bearing specific base sequences complementary to genetic material of different high-risk (hr) HPV types. Synthesis protocol is optimized for scale up production of sensors with high reproducibility. The sensors' surfaces are characterized by scanning electron microscopy (HR-FESEM) and atomic force microscopy (AFM) and their atomic composition is determined by energy dispersive X-ray spectroscopy (EDXS). Oligonucleotide molecules onto nanoporous films block the pores and avoid diffusion of RhB to the liquid phase. Pore opening is produced when specific DNA of HPV is present in the medium, resulting in RhB delivery, that is detected by fluorescence measurements. The sensing assay is optimized for reliable fluorescence signal reading. Nine different sensors are synthesized for specific detection of 14 different hr-HPV types in clinical samples with very high sensitivity (100%) and high selectivity (93-100%), allowing rapid screening of virus infections with very high negative predictive values (100%).
Filiaciones:
Hernández-Montoto A:
The Inter-University Research Institute for Molecular Recognition and Technological Development, Technical University of Valencia, University of Valencia, Camino de Vera s/n, Valencia, 46022, Spain
CIBER Bioengineering, Biomaterials and Nanomedicine, Carlos III Health Institute, Avenida Monforte de Lemos 3-5, Madrid, 28029, Spain
Joint Research Unit in Nanomedicine and Sensors, Health Research Institute Hospital La Fe, Technical University of Valencia, Avenida Fernando Abril Martorell 106, Valencia, 46026, Spain
Aranda MN:
The Inter-University Research Institute for Molecular Recognition and Technological Development, Technical University of Valencia, University of Valencia, Camino de Vera s/n, Valencia, 46022, Spain
Joint Research Unit in Nanomedicine and Sensors, Health Research Institute Hospital La Fe, Technical University of Valencia, Avenida Fernando Abril Martorell 106, Valencia, 46026, Spain
Caballos I:
The Inter-University Research Institute for Molecular Recognition and Technological Development, Technical University of Valencia, University of Valencia, Camino de Vera s/n, Valencia, 46022, Spain
Joint Research Unit in Nanomedicine and Sensors, Health Research Institute Hospital La Fe, Technical University of Valencia, Avenida Fernando Abril Martorell 106, Valencia, 46026, Spain
López-Palacios A:
The Inter-University Research Institute for Molecular Recognition and Technological Development, Technical University of Valencia, University of Valencia, Camino de Vera s/n, Valencia, 46022, Spain
Joint Research Unit in Nanomedicine and Sensors, Health Research Institute Hospital La Fe, Technical University of Valencia, Avenida Fernando Abril Martorell 106, Valencia, 46026, Spain
Tormo-Mas MÁ:
Accredited Research Group on Serious Infection, Health Research Institute Hospital La Fe, Avenida Fernando Abril Martorell 106, Valencia, 46026, Spain
Pemán J:
Accredited Research Group on Serious Infection, Health Research Institute Hospital La Fe, Avenida Fernando Abril Martorell 106, Valencia, 46026, Spain
Microbiology Service, Polytechnic and University Hospital La Fe, Avenida Fernando Abril Martorell 106, Valencia, 46026, Spain
Rodríguez MP:
Pathological Anatomy Service, Polytechnic and University Hospital La Fe, Avenida Fernando Abril Martorell 106, Valencia, 46026, Spain
Picornell C:
Arafarma Group, C/ Fray Gabriel de San Antonio, 6-10, Marchamalo, 19180, Guadalajara, Spain
:
The Inter-University Research Institute for Molecular Recognition and Technological Development, Technical University of Valencia, University of Valencia, Camino de Vera s/n, Valencia, 46022, Spain
CIBER Bioengineering, Biomaterials and Nanomedicine, Carlos III Health Institute, Avenida Monforte de Lemos 3-5, Madrid, 28029, Spain
Joint Research Unit in Nanomedicine and Sensors, Health Research Institute Hospital La Fe, Technical University of Valencia, Avenida Fernando Abril Martorell 106, Valencia, 46026, Spain
UPV-CIPF Joint Research Unit in Mechanisms of Diseases and Nanomedicine, Valencia, Technical University of Valencia, València, 46012, Spain
:
The Inter-University Research Institute for Molecular Recognition and Technological Development, Technical University of Valencia, University of Valencia, Camino de Vera s/n, Valencia, 46022, Spain
CIBER Bioengineering, Biomaterials and Nanomedicine, Carlos III Health Institute, Avenida Monforte de Lemos 3-5, Madrid, 28029, Spain
Joint Research Unit in Nanomedicine and Sensors, Health Research Institute Hospital La Fe, Technical University of Valencia, Avenida Fernando Abril Martorell 106, Valencia, 46026, Spain
UPV-CIPF Joint Research Unit in Mechanisms of Diseases and Nanomedicine, Valencia, Technical University of Valencia, València, 46012, Spain
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