title: Quantitative real-time monitoring of RCA amplification of cancer biomarkers mediated by a flexible ion sensitive platform.
abstract: Ion sensitive field-effect transistors (ISFET) are the basis of radical new sensing approaches. Reliable molecular characterization of specific detection of DNA and/or RNA is vital for disease diagnostics and to follow up alterations in gene expression profiles. Devices and strategies for biomolecular recognition and detection should be developed into reliable and inexpensive platforms. Here, we describe the development of a flexible thin-film sensor for label free gene expression analysis. A charge modulated ISFET based sensor was integrated with real-time DNA/RNA isothermal nucleic acid amplification: Loop-mediated isothermal amplification (LAMP) and Rolling Circle Amplification (RCA) techniques for c-MYC and BCR-ABL1 genes, allowing for the real-time quantification of template. Also, RCA allowed the direct quantification of RNA targets at room temperature, eliminating the requirement for external temperature controllers and overall complexity of the molecular diagnostic approach. This integration between the biological and the sensor/electronic approaches enabled the development of an inexpensive and direct gene expression-profiling platform.
authors.affiliation: UCIBIO, Departamento de Ciências da Vida, Faculdade de Ciências e Tecnologia, Universidade NOVA de Lisboa, Campus de Caparica, 2829-516 Caparica, Portugal; i3N/CENIMAT, Departmento de Ciências de Materiais, Faculdade de Ciências e Tecnologia, Universidade NOVA de Lisboa, Campus de Caparica, 2829-516 Caparica, Portugal.
authors.forename: Bruno
authors.initials: B
authors.lastname: Veigas
authors.affiliation: i3N/CENIMAT, Departmento de Ciências de Materiais, Faculdade de Ciências e Tecnologia, Universidade NOVA de Lisboa, Campus de Caparica, 2829-516 Caparica, Portugal.
authors.forename: Joana
authors.initials: J
authors.lastname: Pinto
authors.affiliation: UCIBIO, Departamento de Ciências da Vida, Faculdade de Ciências e Tecnologia, Universidade NOVA de Lisboa, Campus de Caparica, 2829-516 Caparica, Portugal.
authors.forename: Raquel
authors.initials: R
authors.lastname: Vinhas
authors.affiliation: i3N/CENIMAT, Departmento de Ciências de Materiais, Faculdade de Ciências e Tecnologia, Universidade NOVA de Lisboa, Campus de Caparica, 2829-516 Caparica, Portugal.
authors.forename: Tomás
authors.initials: T
authors.lastname: Calmeiro
authors.affiliation: i3N/CENIMAT, Departmento de Ciências de Materiais, Faculdade de Ciências e Tecnologia, Universidade NOVA de Lisboa, Campus de Caparica, 2829-516 Caparica, Portugal.
authors.forename: Rodrigo
authors.initials: R
authors.lastname: Martins
authors.affiliation: i3N/CENIMAT, Departmento de Ciências de Materiais, Faculdade de Ciências e Tecnologia, Universidade NOVA de Lisboa, Campus de Caparica, 2829-516 Caparica, Portugal.
authors.forename: Elvira
authors.initials: E
authors.lastname: Fortunato
authors.affiliation: UCIBIO, Departamento de Ciências da Vida, Faculdade de Ciências e Tecnologia, Universidade NOVA de Lisboa, Campus de Caparica, 2829-516 Caparica, Portugal. Electronic address: pmvb@fct.unl.pt.
authors.forename: Pedro Viana
authors.initials: PV
authors.lastname: Baptista
mesh_terms: D014408:Biomarkers, Tumor / Q000235:genetics; D015374:Biosensing Techniques / Q000295:instrumentation; D045744:Cell Line, Tumor; D004566:Electrodes; D004867:Equipment Design; D016044:Fusion Proteins, bcr-abl / Q000235:genetics; D020869:Gene Expression Profiling / Q000295:instrumentation*; D016259:Genes, myc; D006801:Humans; D015464:Leukemia, Myelogenous, Chronic, BCR-ABL Positive / Q000175:diagnosis / Q000235:genetics; D009369:Neoplasms / Q000175:diagnosis / Q000235:genetics*; D021141:Nucleic Acid Amplification Techniques / Q000295:instrumentation*
chemical_list: D014408:Biomarkers, Tumor; D016044:Fusion Proteins, bcr-abl
keywords: Cancer; Cmisfet; Field-effect sensors; Gene expression analysis; Rca