Development of a rapid bead-based microfluidic platform for DNA hybridization using single- and multi-mode interactions for probe immobilization

Caneira, Catarina R.F.; Soares, Ruben R. G.; Pinto, Inês F.; Mueller-Landau, Hanna S.; Azevedo, Ana M.; Chu, V.; Conde, J.P.

doi:10.1016/j.snb.2019.01.133

Cited by 19 publications

(11 citation statements)

References 45 publications

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“…Biosensor platforms offer advantages when compared to conventional analytical methods—they are miniaturized and portable, which permits their use as on-site devices, they are also often cheap, simple to use, and do not require any sample preparation. Various sensing technologies were tested for DNA detection, including optical [ 17 ], mass balance [ 18 , 19 ], and quartz crystal microbalance [ 20 , 21 ]. Technological advances have led to the development of miniaturized and sensitive photodetectors termed complementary metal-oxide-semiconductor (CMOS) sensors [ 22 ].…”

Section: Introductionmentioning

confidence: 99%

The Incorporation of Amplified Metal-Enhanced Fluorescence in a CMOS-Based Biosensor Increased the Detection Sensitivity of a DNA Marker of the Pathogenic Fungus Colletotrichum gloeosporioides

2020

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Half of the global agricultural fresh produce is lost, mainly because of rots that are caused by various pathogenic fungi. In this study, a complementary metal-oxide-semiconductor (CMOS)-based biosensor was developed, which integrates specific DNA strands that allow the detection of enoyl-CoA-hydratase/isomerase, which is a quiescent marker of Colletotrichum gloeosporioides fungi. The developed biosensor mechanism is based on the metal-enhanced fluorescence (MEF) phenomenon, which is amplified by depositing silver onto a glass surface. A surface DNA strand is then immobilized on the surface, and in the presence of the target mRNA within the sample, the reporter DNA strand that is linked to horseradish peroxidase (HRP) enzyme will also bind to it. The light signal that is later produced from the HRP enzyme and its substrate is enhanced and detected by the coupled CMOS sensor. Several parameters that affect the silver-deposition procedure were examined, including silver solution temperature and volume, heating mode, and the tank material. Moreover, the effect of blocking treatment (skim milk or bovine serum albumin (BSA)) on the silver-layer stability and nonspecific DNA absorption was tested. Most importantly, the effect of the deposition reaction duration on the silver-layer formation and the MEF amplification was also investigated. In the study findings a preferred silver-deposition reaction duration was identified as 5–8 min, which increased the deposition of silver on the glass surface up to 13-times, and also resulted in the amplification of the MEF phenomenon with a maximum light signal of 50 relative light units (RLU). It was found that MEF can be amplified by a customized silver-deposition procedure that results in increased detection sensitivity. The implementation of the improved conditions increased the biosensor sensitivity to 3.3 nM (4500 RLU) with a higher detected light signal as compared to the initial protocol (400 RLU). Moreover, the light signal was amplified 18.75-, 11.11-, 5.5-, 11.25-, and 3.75-times in the improved protocol for all the tested concentrations of the target DNA strand of 1000, 100, 10, 3.3, and 2 nM, respectively. The developed biosensor system may allow the detection of the pathogenic fungus in postharvest produce and determine its pathogenicity state.

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Section: Introductionmentioning

confidence: 99%

The Incorporation of Amplified Metal-Enhanced Fluorescence in a CMOS-Based Biosensor Increased the Detection Sensitivity of a DNA Marker of the Pathogenic Fungus Colletotrichum gloeosporioides

2020

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“…The sensitivity was maximized by using QDs as optical labels, and a low LOD of approximately 9.5 pM was achieved. 9 Ankireddy et al designed a QD-bead-DNA conjugate probe integrated into two channels of a PDMS microfluidic device for the detection of Met gene-related cancer. This probe was able to distinguish wild-type from mutant DNA at a low DNA concentration of 1 nM.…”

Section: Detection Of Nucleic Acidsmentioning

confidence: 99%

“…Thus far, the microfluidic technique is able to integrate multiple functions including fluid manipulation, cell capture, culture, sorting and detection as a single device for pathogenic cell detection, gene and protein analyses, cancer biology and diagnoses. [7][8][9] In addition to QDs as fluorescent biosensors for detection on microfluidic chips, in vivo fluorescence imaging (FI) is another novel application that has rapidly developed over the last decade. Cell activity and gene behavior in living organisms can be monitored by in vivo FI for analyzing biological processes such as tumor growth and metastasis, stem cell therapy and specific gene expression.…”

Section: Introductionmentioning

confidence: 99%

Micro-/nano-fluidic devices and in vivo fluorescence imaging based on quantum dots for cytologic diagnosis

2022

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“…It can be used for the direct recognition of specific target sequences, with applications in molecular biology and molecular diagnostics, since particular diseases can be identified based on the identification of specific nucleic acid sequences. Examples are the detection of methicillin-resistant Staphylococcus aureus (MRSA) [ 4 ], mi-RNA sequences related to tumor or cardiovascular diseases [ 5 , 6 ], or coronavirus-related sequences [ 7 , 8 ]. In this respect, in response to the pandemic COVID-19 emergency, the selective recognition of SARS-CoV-2 sequences has been recently demonstrated using a thermoplasmonic approach based on DNA hybridization [ 9 ].…”

Section: Introductionmentioning

confidence: 99%

Morphological and Mechanical Characterization of DNA SAMs Combining Nanolithography with AFM and Optical Methods

et al. 2020

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The morphological and mechanical properties of thiolated ssDNA films self-assembled at different ionic strength on flat gold surfaces have been investigated using Atomic Force Microscopy (AFM). AFM nanoshaving experiments, performed in hard tapping mode, allowed selectively removing molecules from micro-sized regions. To image the shaved areas, in addition to the soft contact mode, we explored the use of the Quantitative Imaging (QI) mode. QI is a less perturbative imaging mode that allows obtaining quantitative information on both sample topography and mechanical properties. AFM analysis showed that DNA SAMs assembled at high ionic strength are thicker and less deformable than films prepared at low ionic strength. In the case of thicker films, the difference between film and substrate Young’s moduli could be assessed from the analysis of QI data. The AFM finding of thicker and denser films was confirmed by X-Ray Photoelectron Spectroscopy (XPS) and Spectroscopic Ellipsometry (SE) analysis. SE data allowed detecting the DNA UV absorption on dense monomolecular films. Moreover, feeding the SE analysis with the thickness data obtained by AFM, we could estimate the refractive index of dense DNA films.

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Development of a rapid bead-based microfluidic platform for DNA hybridization using single- and multi-mode interactions for probe immobilization

Cited by 19 publications

References 45 publications

The Incorporation of Amplified Metal-Enhanced Fluorescence in a CMOS-Based Biosensor Increased the Detection Sensitivity of a DNA Marker of the Pathogenic Fungus Colletotrichum gloeosporioides

The Incorporation of Amplified Metal-Enhanced Fluorescence in a CMOS-Based Biosensor Increased the Detection Sensitivity of a DNA Marker of the Pathogenic Fungus Colletotrichum gloeosporioides

Micro-/nano-fluidic devices and in vivo fluorescence imaging based on quantum dots for cytologic diagnosis

Morphological and Mechanical Characterization of DNA SAMs Combining Nanolithography with AFM and Optical Methods

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