Comparison of Different Strategies for the Development of Highly Sensitive Electrochemical Nucleic Acid Biosensors Using Neither Nanomaterials nor Nucleic Acid Amplification
| Autor: | Víctor Ruiz-Valdepeñas Montiel, Rebeca M. Torrente-Rodríguez, Eva Vargas, José M. Pingarrón, Susana Campuzano, A. Julio Reviejo, Eloy Povedano, María Pedrero |
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| Rok vydání: | 2018 |
| Předmět: |
Bioengineering
Biosensing Techniques 02 engineering and technology Computational biology 01 natural sciences Antibodies Nanomaterials chemistry.chemical_compound Nucleic Acids Instrumentation Fluid Flow and Transfer Processes Staining and Labeling Process Chemistry and Technology 010401 analytical chemistry Nucleic Acid Hybridization RNA Electrochemical Techniques Equipment Design 021001 nanoscience & nanotechnology 0104 chemical sciences Highly sensitive chemistry Nucleic acid 0210 nano-technology Biosensor DNA |
| Zdroj: | ACS Sensors. 3:211-221 |
| ISSN: | 2379-3694 |
| Popis: | Currently, electrochemical nucleic acid-based biosensing methodologies involving hybridization assays, specific recognition of RNA/DNA and RNA/RNA duplexes, and amplification systems provide an attractive alternative to conventional quantification strategies for the routine determination of relevant nucleic acids at different settings. A particularly relevant objective in the development of such nucleic acid biosensors is the design of as many as possible affordable, quick, and simple methods while keeping the required sensitivity. With this aim in mind, this work reports, for the first time, a thorough comparison between 11 methodologies that involve different assay formats and labeling strategies for targeting the same DNA. The assayed approaches use conventional sandwich and competitive hybridization assays, direct hybridization coupled to bioreceptors with affinity for RNA/DNA duplexes, multienzyme labeling bioreagents, and DNA concatamers. All of them have been implemented on the surface of magnetic beads (MBs) and involve amperometric transduction at screen-printed carbon electrodes (SPCEs). The influence of the formed duplex length and of the labeling strategy have also been evaluated. Results demonstrate that these strategies can provide very sensitive methods without the need for using nanomaterials or polymerase chain reaction (PCR). In addition, the sensitivity can be tailored within several orders of magnitude simply by varying the bioassay format, hybrid length or labeling strategy. This comparative study allowed us to conclude that the use of strategies involving longer hybrids, the use of antibodies with specificity for RNA/DNA heteroduplexes and labeling with bacterial antibody binding proteins conjugated with multiple enzyme molecules, provides the best sensitivity. |
| Databáze: | OpenAIRE |
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