4.7 Article

Upconversion-Linked Immunoassay for the Diagnosis of Honeybee Disease American Foulbrood

Publisher

IEEE-INST ELECTRICAL ELECTRONICS ENGINEERS INC
DOI: 10.1109/JSTQE.2021.3049689

Keywords

American foulbrood; bacterial pathogens; Paenibacillus larvae; photon-upconversion nanoparticle; upconversion-linked immunosorbent assay

Funding

  1. Ministry of Education, Youth and Sports of the Czech Republic (MEYS CR) under the Project INTER-ACTION [LTAB19011]
  2. CFNanobiotechnology of CIISB, Instruct-CZ Centre by MEYS CR [LM2018127]
  3. CF Cryo-Electron Microscopy and Tomography of CIISB, Instruct-CZ Centre by MEYS CR [LM2018127]
  4. CzechNanoLab Project - MEYS CR [LM2018110]
  5. Czech Science Foundation [18-03367Y]
  6. Institute of Analytical Chemistry of the Czech Academy of Sciences [RVO 68081715]
  7. German Research Foundation [DFG: GO 1968/5-1, GO 1968/7-1]
  8. MEYS CR under the Project CEITEC 2020 [LQ1601]
  9. Bavarian-Czech Academic Agency (BTHA)

Ask authors/readers for more resources

By developing a ULISA method with a specific antibody for Paenibacillus larvae, this study successfully detected bacterial loads present in honeybee samples with low cross-reactivity and improved detection limits, providing a valuable tool for AFB diagnosis and monitoring.
American foulbrood (AFB) caused by the bacterium Paenibacillus larvae is the most destructive disease of the honeybee brood. Therefore, rapid and sensitive detection methods are required to limit spreading of this pathogen, which has a major impact on agriculture and biodiversity. While P. larvae is typically detected by microbial cultivation or polymerase chain reaction, antibody-based detection represents a viable alternative. Here, we prepared an antibody specific for P. larvae and used it for the development of an upconversion-linked immunosorbent assay (ULISA). Photon-upconversion nanoparticles (UCNP) were conjugated to streptavidin via a PEG-linker using copper-catalyzed click chemistry to replace the enzyme label in conventional enzyme-linked immunosorbent assay (ELISA). The ULISA showed low cross-reactivity and provided a limit of detection of 2.9 x 10(3) CFU/mL, representing a 22-fold improvement compared to the ELISA. This level is within the bacterial loads present in honeybee larvae during an AFB infection. The assay was successfully applied to the analysis of spiked samples of bees, larvae, and hive debris.

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