title: Ion sensing (EIS) real-time quantitative monitorization of isothermal DNA amplification.
abstract: Field-effect-based devices are becoming a basic structural element in a new generation of microbiosensors. Reliable molecular characterization of DNA and/or RNA is of paramount importance for disease diagnostics and to follow up alterations in gene expression profiles. The use of such devices for point-of-need diagnostics has been hindered by the need of standard or real-time PCR amplification procedures. The present work focuses on the development of a tantalum pentoxide (Ta2O5) based sensor for the real-time label free detection of DNA amplification via loop mediated isothermal amplification (LAMP) allowing for quantitative analysis of the cMYC proto-oncogene. The strategy based on the field effect sensor was tested within a range of 1 × 10(8)-10(11) copies of target DNA, and a linear relationship between the log copy number of the initial template DNA and threshold time was observed allowing for a semi-quantitative analysis of DNA template. The concept offers many of the advantages of isothermal quantitative real-time DNA amplification in a label free approach and may pave the way to point-of-care quantitative molecular analysis focused on ease of use and low cost.
authors.affiliation: CIGMH, Departamento de Ciências da Vida, Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa, Campus de Caparica, 2829-516 Caparica, Portugal; CENIMAT/I3N, Departamento de Ciência dos Materiais, Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa and CEMOP-UNINOVA, Campus de Caparica, 2829-516 Caparica, Portugal.
authors.forename: Bruno
authors.initials: B
authors.lastname: Veigas
authors.forename: Rita
authors.initials: R
authors.lastname: Branquinho
authors.forename: Joana V
authors.initials: JV
authors.lastname: Pinto
authors.forename: Pawel J
authors.initials: PJ
authors.lastname: Wojcik
authors.forename: Rodrigo
authors.initials: R
authors.lastname: Martins
authors.forename: Elvira
authors.initials: E
authors.lastname: Fortunato
authors.forename: Pedro V
authors.initials: PV
authors.lastname: Baptista
mesh_terms: D015374:Biosensing Techniques / Q000379:methods*; D004247:DNA / Q000302:isolation & purification*; D021141:Nucleic Acid Amplification Techniques; D010087:Oxides / Q000737:chemistry; D012313:RNA / Q000302:isolation & purification*; D013635:Tantalum / Q000737:chemistry
chemical_list: D010087:Oxides; D012313:RNA; D013635:Tantalum; D004247:DNA; C078151:tantalum oxide
keywords: Cancer; DNA; EIS; Ion sensitive field effect sensors; Isothermal amplification; LAMP; Label free; Real time; Tantalum pentoxide; c-Myc; qRT-PCR