| 1. |
- Acero Sanchez, Josep Ll., et al.
(författare)
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Electrochemical Genetic Profiling of Single Cancer Cells
- 2017
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Ingår i: Analytical Chemistry. - : American Chemical Society (ACS). - 0003-2700 .- 1520-6882. ; 89:6, s. 3378-3385
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Tidskriftsartikel (refereegranskat)abstract
- Recent understandings in the development and spread of cancer have led to the realization of novel single cell analysis platforms focused on circulating tumor cells (CTCs). A simple, rapid, and inexpensive analytical platform capable of providing genetic information on these rare cells is highly desirable to support clinicians and researchers alike to either support the selection or adjustment of therapy or provide fundamental insights into cell function and cancer progression mechanisms. We report on the genetic profiling of single cancer cells, exploiting a combination of multiplex ligation-dependent probe amplification (MLPA) and electrochemical detection. Cells were isolated using laser capture and lysed, and the mRNA was extracted and transcribed into DNA. Seven markers were amplified by MLPA, which allows for the simultaneous amplification of multiple targets with a single primer pair, using MLPA probes containing unique barcode sequences. Capture probes complementary to each of these barcode sequences were immobilized on a printed circuit board (PCB) manufactured electrode array and exposed to single-stranded MLPA products and subsequently to a single stranded DNA reporter probe bearing a HRP molecule, followed by substrate addition and fast electrochemical pulse amperometric detection. We present asimple, rapid, flexible, and inexpensive approach for the simultaneous quantification of multiple breast cancer related mRNA markers, with single tumor cell sensitivity.
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| 2. |
- Debela, A. M., et al.
(författare)
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Functionalized deoxynucleotides and DNA primers for electrochemical diagnostics of disease predispostions
- 2017
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Ingår i: ECS Transactions. - : Electrochemical Society Inc.. - 1938-6737 .- 1938-5862. - 9781607685395 ; , s. 1873-1883, s. 1873-1883
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Konferensbidrag (refereegranskat)abstract
- Redox labeled DNAs are of increasing interest for the fabrication of next generation molecular tools. In the present work we are investigating the use of various redox labeled dNTPs, ddNTPs and DNA primers for use in detection of diseases. We have reported the use of Polyoxometalate (POM) labeled DNA primers and dNTPs for use in PCR and subsequently used for direct electrochemical detection of PCR products. The use of POM labeled DNAs in PCR enabled us to check the compatibility with polymerases and PCR incorporability of the modified DNAs. Furthermore we have investigated the solid-phase array based primer extension (é-PEX) with redox labelled ddNTPs (ferrocene (Fc), anthraquinone (AQ) phenothiazine (PTZ) and methylene blue (MB)) to prove the strategy of detection of single nucleotide polymorphisms using the labeled ddNTPs. This strategy will allow the development of cost-effective, rapid and user-friendly platform for the screening of known and unknown genetic mutations.
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| 3. |
- Hernandez, Frank Jeyson, et al.
(författare)
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Label free optical sensor for Avidin based on single gold nanoparticles functionalized with aptamers
- 2009
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Ingår i: Journal of Biophotonics. - : Wiley-VCH Verlagsgesellschaft. - 1864-063X .- 1864-0648. ; 2:4, s. 227-231
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Tidskriftsartikel (refereegranskat)abstract
- Optical spectroscopy of a single gold nanoparticle, functionalized with an aptamer, is used to sense the specific binding of avidin. Herewith, the field of single noble metal nanoparticle biosensors is extended to the important field of aptamer based assays. The sensitivity of this initial, but not yet optimized apta-nano-sensor is in the range of 20 nM. Due to its nanoscopic size, this single nanoparticle based apta-sensor may be used in nanoscopic volumes such as in array type assays or even inside cells.
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| 4. |
- Toldrà Filella, Anna, et al.
(författare)
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Detecting harmful algal blooms with nucleic acid amplification-based biotechnological tools
- 2020
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Ingår i: Science of the Total Environment. - : Elsevier BV. - 0048-9697 .- 1879-1026. ; 749
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Forskningsöversikt (refereegranskat)abstract
- Harmful algal blooms (HABs) represent a growing threat to aquatic ecosystems and humans. Effective HAB management and mitigation efforts strongly rely on the availability of timely and in-situ tools for the detection of microalgae. In this sense, nudeic acid-based (molecular) methods are being considered for the unequivocal identification of miaoalgae as an attractive alternative to the currently used time-consuming and laboratory-based light microscopy techniques. This review provides an overview of the progress made on new molecular biotechnological tools for microalgal detection, particularly focusing on those that combine a nudeic acid (DNA or RNA) amplification step with detection. Different types of amplification processes (thermal and isothermal) and detection formats (e.g. microarrays, biosensors, lateral flows) are presented, and a comprehensive overview of their advantages and limitations is provided Although isothermal techniques are an attractive alternative to thermal amplification to reach in-situ analysis, further development is still required. Finally, current challenges, critical steps and future directions of the whole analysis process ( from sample procurement to in-situ implementation) are described.
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| 5. |
- Torrens, Mabel, et al.
(författare)
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Amperometric detection of Francisella tularensis genomic sequence on Zn-mediated diazonium modified substrates
- 2015
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Ingår i: Electrochemistry communications. - : Elsevier. - 1388-2481 .- 1873-1902. ; 53
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Tidskriftsartikel (refereegranskat)abstract
- A simple, rapid and potentiostat-free method for grafting of close to monolayers of diazonium salts to anchor DNA probes for application in highly sensitive electrochemical genosensing is presented. Zn-mediated grafting is demonstrated to produce a highly organised surface facilitating enhanced sensitivity and a notably reduced detection limit as compared to electrochemical grafting. (C) 2015 Elsevier B.V. All rights reserved.
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