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Amplified plasmonic and microfluidic setup for DNA monitoring

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Abstract

Plasmonic nanosensors for label-free detection of DNA require excellent sensing resolution, which is crucial when monitoring short DNA sequences, as these induce tiny peak shifts, compared to large biomolecules. We report a versatile and simple strategy for plasmonic sensor signal enhancement by assembling multiple (four) plasmonic sensors in series. This approach provided a fourfold signal enhancement, increased signal-to-noise ratio, and improved sensitivity for DNA detection. The response of multiple sensors based on AuNSpheres was also compared with  AuNRods, the latter showing better sensing resolution. The amplification system based on AuNR was integrated into  a microfluidic sequential injection platform and applied to the monitoring of DNA, specifically from environmental invasive species—zebra mussels. DNA from zebra mussels was log concentration-dependent from 1 to 1 × 106 pM, reaching a detection limit of 2.0 pM. In situ tests were also successfully applied to real samples, within less than 45 min, using DNA extracted from zebra mussel meat. The plasmonic nanosensors’ signal can be used as a binary output (yes/no) to assess the presence of those invasive species. Even though these genosensors were applied to the monitoring of DNA in environmental samples, they potentially offer advantage in a wide range of fields, such as disease diagnostics.

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Funding

This work was supported by project Nanotechnology Based Functional Solutions (NORTE-01-0145-FEDER-000019), supported by Norte Portugal Regional Operational Programme (NORTE2020), under the PORTUGAL 2020 Partnership Agreement, through the European Regional Development Fund, and by the Partnership Agreement with Confederación Hidrográfica del Guadalquivir (Spain) for the development of a system of early detection of the zebra mussel through analysis of environmental DNA.

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Authors and Affiliations

  1. Food Quality and Safety Research Group, International Iberian Nanotechnology Laboratory (INL), Avenida Mestre José Veiga, 4715-310, Braga, Portugal

    J. Rafaela L. Guerreiro, Andrey Ipatov, Joana Carvalho & Marta Prado

  2. IRTA, Ctra. Poble Nou Km 5.5 Sant Carles de la Ràpita, 43540, Tarragona, Spain

    Anna Toldrà

Authors
  1. J. Rafaela L. Guerreiro
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  2. Andrey Ipatov
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  3. Joana Carvalho
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  4. Anna Toldrà
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  5. Marta Prado
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Contributions

Experiments were conducted by J.R. Guerreiro and A. Toldrà. Flow cell design and fabrication, as well as microfluidic assembly and software development, were conducted by A. Ipatov. Zebra mussel-specific DNA sequence identification and extraction/preparation of real samples were conducted by J. Carvalho. Conceptualization and method development were designed by J.R. Guerreiro and M. Prado. Writing—original draft preparation, was conducted by J.R. Guerreiro. M. Prado participated on project administration and funding acquisition. Writing—reviewing and editing, was contributed by J.R. Guerreiro, A. Ipatov, J. Carvalho, A. Toldrà, and M. Prado.

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Correspondence to J. Rafaela L. Guerreiro.

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Guerreiro, J.R.L., Ipatov, A., Carvalho, J. et al. Amplified plasmonic and microfluidic setup for DNA monitoring. Microchim Acta 188, 326 (2021). https://doi.org/10.1007/s00604-021-04983-y

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