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A dual-targeting nanobiosensor for Gender Determination applying Signal Amplification Methods and integrating Fluorometric Gold and Silver Nanoclusters

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Abstract

A dual-targeting nanobiosensor has been developed for the simultaneous detection of AMELX and AMELY genes based on the different fluorescence signals emitted from gold and silver nanoclusters, AuNCs and AgNCs respectively. In our design, both catalytic hairpin assembly (CHA) and hybridization chain reaction (HCR) have been used as isothermal, enzyme-free and simple methods for signal’s amplification. The working principle is based on the initiation of a cascade of CHA-HCR reactions when AMELX is present, in which AuNCs, synthesized on the third hairpin, are aggregated on the surface of the dsDNA product, performing the phenomenon of aggregation induced emission (AIE) and enhancing their fluorescence signal. On the other hand, the presence of the second target, AMELY, is responsible for the enhancement of the fluorescence signal corresponding to AgNCs by the same phenomenon, via hybridizing to the free end of the dsDNA formed and at the same time to the probe of silver nanoclusters fixing it closer to the surface of the dsDNA product. Such a unique design has the merits of being simple, inexpensive, specific and stable and presents rapid results. The detection limits of this assay for AMELX and AMELY are as low as 3.16 fM and 23.6 fM respectively. Moreover, this platform showed great performance in real samples. The design has great promise for the application of dual-targeting nanobiosensors to other biomarkers.

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Acknowledgements

Gratefully, this study was financially supported by the research Council of University of Tehran.

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Authors and Affiliations

  1. Nanobiosensors Lab, Department of Life Science Engineering, Faculty of New Sciences and Technologies, University of Tehran, P.O. Box 14399-56191, Tehran, Iran

    Fatima Bazzi, Morteza Hosseini & Bahman Ebrahimi-Hoseinzadeh

  2. Department of Chemistry, College of Science, Sultan Qaboos University, Box 36, Al-Khod 123, Sultan Qaboos, Oman

    Haider A. J. Al Lawati

  3. School of Chemistry, Faculty of Science, University of Tehran, P.O. Box 14176-14411, Tehran, Iran

    Mohammad Reza Ganjali

Authors
  1. Fatima Bazzi
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  2. Morteza Hosseini
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  3. Bahman Ebrahimi-Hoseinzadeh
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  4. Haider A. J. Al Lawati
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  5. Mohammad Reza Ganjali
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Contributions

Fatima Bazzi: Formal analysis, visualization, data curation, writing-original, draft, review and editing., Morteza Hosseini: Conceptualization, methodology, designing the analysis, visualization, writing-review and editing. Bahman Ebrahimi-Hoseinzadeh: Project administration, visualization, designing the analysis, Haider A.J. Al Lawati: writing-review and editing, Project consultant and Mohammad Reza Ganjali: Writing-review and editing.

Corresponding authors

Correspondence to Morteza Hosseini or Bahman Ebrahimi-Hoseinzadeh.

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The authors have no financial or non-financial conflicts of interest to disclose.

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AMELX, Amelogenin gene X; AMELY, Amelogenin gene Y; AuNCs, Gold nanoclusters; AgNCs, silver nanoclusters; CHA, catalytic hairpin assembly; HCR, hybridization chain reaction; AIE, aggregation induced emission.

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Bazzi, F., Hosseini, M., Ebrahimi-Hoseinzadeh, B. et al. A dual-targeting nanobiosensor for Gender Determination applying Signal Amplification Methods and integrating Fluorometric Gold and Silver Nanoclusters. Microchim Acta 190, 368 (2023). https://doi.org/10.1007/s00604-023-05947-0

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