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An Integrated System to Remotely Trigger Intracellular Signal Transduction by Upconversion Nanoparticle-mediated Kinase Photoactivation
Published on: August 30, 2017
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Click-conjugated photon-upconversion nanoparticles in an immunoassay for honeybee pathogen Melissococcus plutonius
Veronika Poláchová1, Matěj Pastucha, Zuzana Mikušová
1CEITEC MU, Masaryk University, Kamenice 5, 625 00 Brno, Czech Republic. farka@mail.muni.cz.
Nanoscale
|April 16, 2019
Summary
A new upconversion-linked immunosorbent assay (ULISA) offers ultrasensitive detection of European foulbrood (EFB) in honeybees. This advanced method significantly improves early diagnosis of Melissococcus plutonius infection, outperforming traditional ELISA.
Area of Science:
- Nanotechnology-based biosensing
- Apiculture and bee health
- Infectious disease diagnostics
Background:
- European foulbrood (EFB), caused by Melissococcus plutonius, severely impacts honeybee colonies.
- Conventional enzyme-linked immunosorbent assay (ELISA) lacks the sensitivity for early EFB detection.
- Photon-upconversion nanoparticles (UCNPs) offer ultrasensitive detection by minimizing optical background interference.
Purpose of the Study:
- To develop a highly sensitive assay for early detection of Melissococcus plutonius.
- To conjugate UCNPs with streptavidin for enhanced immunoassay performance.
- To evaluate the efficacy of the developed assay in real-world samples.
Main Methods:
- Surface modification of UCNPs with alkyne-BSA via copper-free click chemistry for stable conjugation.
- Development of a sandwich upconversion-linked immunosorbent assay (ULISA) using anti-Melissococcus antibodies.
- Detection of M. plutonius in spiked bee samples, larvae, and hive debris.
Main Results:
- The ULISA achieved a limit of detection (LOD) of 340 CFU mL−1, 400 times more sensitive than ELISA.
- The assay demonstrated a wide working range, up to 10^9 CFU mL−1.
- Successful detection of M. plutonius in various spiked bee-related samples confirmed practical applicability.
Conclusions:
- The UCNP-based ULISA provides a highly sensitive and effective tool for early EFB diagnosis.
- This assay has the potential to prevent the spread of EFB and reduce honeybee colony losses.
- The developed conjugation chemistry and assay format are promising for other bacterial detection applications.
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