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Aptamer based fluorometric acetamiprid assay using three kinds of nanoparticles for powerful signal amplification

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Abstract

The authors describe an aptamer-based fluorometric assay for the insecticide acetamiprid. It is based on target-induced release of the fluorescein-labeled complementary strand of the aptamer (CS) from the aptamer/CS conjugate (dsDNA). Three kinds of nanoparticles with opposite effects on the fluorophore (FAM) were used. These include gold nanoparticles (AuNPs), single-walled carbon nanotubes (SWNTs) and silica nanoparticles (SiNPs) coated with streptavidin. In the presence of acetamiprid, FAM-labeled CS is released from the dsDNA-modified SNP-streptavidin complex and accumulates in the supernatant (phase I) after centrifugation. Fluorescence intensity decreases on addition of the supernatant to the SWNTs and AuNPs because they act as quenchers (phase II). In the absence of acetamiprid, the dsDNA-modified SiNP-streptavidin complex remains intact and no labeled CS is present in the supernatant containing the AuNPs and SWNTs. So, the relative fluorescence intensity is quite low. The assay is highly selective for acetamiprid and has a limit of detection (LOD) as low as 127 pM. The method was successfully applied to the determination of acetamiprid in spiked serum and water where it gave LODs of 198 and 130 pM, respectively.

In the absence of acetamiprid, the dsDNA-modified silica nanoparticle (SiNP)-streptavidin conjugate remains intact, leading to a very weak relative fluorescence intensity. In the presence of target, the dsDNA-modified SiNP-streptavidin complex is disassembled and FAM-labeled CS is released from the aptamer (Apt), resulting in a very strong relative fluorescence intensity.

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Acknowledgments

Financial support of this study was provided by Mashhad University of Medical Sciences.

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Authors and Affiliations

  1. Pharmaceutical Research Center, Mashhad University of Medical Sciences, Mashhad, Iran

    Khalil Abnous & Mona Alibolandi

  2. Academic Center For Education, Culture and Research (ACECR)-Mashhad Branch, Mashhad, Iran

    Khalil Abnous & Parirokh Lavaee

  3. Nanotechnology Research Center, Mashhad University of Medical Sciences, Mashhad, Iran

    Noor Mohammad Danesh & Mohammad Ramezani

  4. Research Institute of Sciences and New Technology, Mashhad, Iran

    Noor Mohammad Danesh

  5. Targeted Drug Delivery Research Center, Mashhad University of Medical Sciences, Mashhad, Iran

    Seyed Mohammad Taghdisi

  6. Department of Pharmaceutical Biotechnology, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran

    Seyed Mohammad Taghdisi

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  1. Khalil Abnous
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  2. Noor Mohammad Danesh
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  6. Seyed Mohammad Taghdisi
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Correspondence to Seyed Mohammad Taghdisi.

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Khalil Abnous and Noor Mohammad Danesh contributed equally to this work.

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Abnous, K., Danesh, N.M., Ramezani, M. et al. Aptamer based fluorometric acetamiprid assay using three kinds of nanoparticles for powerful signal amplification. Microchim Acta 184, 81–90 (2017). https://doi.org/10.1007/s00604-016-1992-3

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  • DOI: https://doi.org/10.1007/s00604-016-1992-3

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