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Ultrasensitive detection of Staphylococcus aureus using a non-fluorescent cDNA-grafted dark BBQ®-650 chromophore integrated hydrophilic upconversion nanoparticles/aptamer system

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Abstract

A highly structured fluorometric bioassay has been proposed for screening Staphylococcus aureus (S. aureus). The study exploits (i) the spectral attributes of the hexagonal NaYF4:Yb,Er upconversion nanoparticle (UCNP)-coated 3-aminopropyl)triethoxysilane; (ii) the intrinsic non-fluorescent quenching features of the highly stable dark blackberry (BBQ®-650) receptor; (iii) the aptamer (Apt-) biorecognition and binding affinity, and (iv) the complementary DNA hybridizer-linkage efficacy. The principle relied on the excited state energy transfer between the donor Apt-labeled NH2-UCNPs at the 3′ end, and cDNA-grafted BBQ®-650 at the 5′ end, as the effective receptors. The donor moieties in proximity (< 10.0 nm) trigger hybridization with the cDNA-grafted dark BBQ®-650, as the receptors of energy from the 2F5/2 level of Yb3+ ions to initiate the Förster resonance energy transfer pathway. This was confirmed by the decline in the excited-state lifetimes from 223.52 μs (τ1) to 179.26 μs (τ2). The existence of the target S. aureus in the bioassay attracts the Apt- resulting in the detachment of the acceptor, and disintegration of the complex configuration via conformation reversal. The re-activated fluorescence monitored at λex/em = 980/652 nm, as a function of the logarithmic concentration of S. aureus (42 to 4.2 × 108 CFU mL−1), yielded an ultra-low detection response of 2.0 CFU mL−1. The bioassay screening of S. aureus in real samples revealed satisfactory recoveries (92.44–107.82%) and validation results (p > 0.05). Hence, the comprehensive Apt-labeled NH2-UCNPs-cDNA-grafted dark BBQ®-650 bioassay offered fast and precise S. aureus screening in food and environmental settings.

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Funding

The authors received financial support from “entrepreneurship and innovation project of Jiangsu province, China (No: JSSCBS20210930).”

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Authors and Affiliations

  1. College of Food and Biological Engineering, Jimei University, Xiamen, 361021, People’s Republic of China

    Waqas Ahmad & Quansheng Chen

  2. School of Food and Biological Engineering, Jiangsu University, Zhenjiang, 212013, People’s Republic of China

    Waqas Ahmad, Li Wang, Muhammad Zareef & Quansheng Chen

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Ahmad, W., Wang, L., Zareef, M. et al. Ultrasensitive detection of Staphylococcus aureus using a non-fluorescent cDNA-grafted dark BBQ®-650 chromophore integrated hydrophilic upconversion nanoparticles/aptamer system. Microchim Acta 190, 250 (2023). https://doi.org/10.1007/s00604-023-05823-x

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