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 x 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.
Original languageEnglish
Pages (from-to)50-55
Number of pages6
JournalBiosensors & Bioelectronics
Volume52
Issue numberNA
DOIs
Publication statusPublished - 15 Feb 2014

Fingerprint

Ions
DNA
Point-of-Care Systems
Equipment and Supplies
Proto-Oncogenes
Transcriptome
Real-Time Polymerase Chain Reaction
RNA
Costs and Cost Analysis

Keywords

  • DNA
  • LAMP
  • Isothermal amplification
  • EIS
  • Ion sensitive field effect sensors
  • Tantalum pentoxide
  • qRT-PCR
  • Real time
  • c-Myc
  • Cancer
  • Label free

Cite this

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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 x 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.",
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author = "Bruno Veigas and Rita Branquinho and {Vaz Pinto}, Joana and Wojcik, {Pawel Jerzy} and Martins, {Rodrigo Ferr{\~a}o de Paiva} and Fortunato, {Elvira Maria Correia} and Baptista, {Pedro Miguel Ribeiro Viana}",
note = "Sem PDF. Fundacao para a Ciencia e a Tecnologia (FCT-MCTES) (PEst C/CTM/LA0025/2013-14, LA25-2013-2014; PEst-OE/SAU/UI0009/2011; PTDC/BBB-NAN/1812/2012; PTDC/QUI-QUI/112597/2009; PTDC/CTM-NAN/109877/2009; Blood-FET-PTDC/SAU-BEB/098125/2009; Multinox-EXCL/CTM-NAN/0201/2012) ERC (228144) SFRH/BD/78970/2011 SFRH/BD/45224/2008",
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month = "2",
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doi = "10.1016/j.bios.2013.08.029",
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journal = "Biosensors & Bioelectronics",
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publisher = "Elsevier Science B.V., Amsterdam.",
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TY - JOUR

T1 - Ion sensing (EIS) real-time quantitative monitorization of isothermal DNA amplification

AU - Veigas, Bruno

AU - Branquinho, Rita

AU - Vaz Pinto, Joana

AU - Wojcik, Pawel Jerzy

AU - Martins, Rodrigo Ferrão de Paiva

AU - Fortunato, Elvira Maria Correia

AU - Baptista, Pedro Miguel Ribeiro Viana

N1 - Sem PDF. Fundacao para a Ciencia e a Tecnologia (FCT-MCTES) (PEst C/CTM/LA0025/2013-14, LA25-2013-2014; PEst-OE/SAU/UI0009/2011; PTDC/BBB-NAN/1812/2012; PTDC/QUI-QUI/112597/2009; PTDC/CTM-NAN/109877/2009; Blood-FET-PTDC/SAU-BEB/098125/2009; Multinox-EXCL/CTM-NAN/0201/2012) ERC (228144) SFRH/BD/78970/2011 SFRH/BD/45224/2008

PY - 2014/2/15

Y1 - 2014/2/15

N2 - 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 x 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.

AB - 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 x 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.

KW - EIS

KW - Label free

KW - DNA

KW - Cancer

KW - Tantalum pentoxide

KW - qRT-PCR

KW - LAMP

KW - Isothermal amplification

KW - Real time

KW - c-Myc

KW - Ion sensitive field effect sensors

KW - DNA

KW - LAMP

KW - Isothermal amplification

KW - EIS

KW - Ion sensitive field effect sensors

KW - Tantalum pentoxide

KW - qRT-PCR

KW - Real time

KW - c-Myc

KW - Cancer

KW - Label free

U2 - 10.1016/j.bios.2013.08.029

DO - 10.1016/j.bios.2013.08.029

M3 - Article

VL - 52

SP - 50

EP - 55

JO - Biosensors & Bioelectronics

JF - Biosensors & Bioelectronics

SN - 0956-5663

IS - NA

ER -