Immunosensing of the honeybee bacterial pathogen Melissococcus plutonius

PASTUCHA, Matěj, Zdeněk FARKA, Veronika POLÁCHOVÁ, Zuzana MIKUŠOVÁ, Radka OBOŘILOVÁ and Petr SKLÁDAL. Immunosensing of the honeybee bacterial pathogen Melissococcus plutonius. In Joint Retreat 2019. 2019. ISBN 978-80-210-9300-3.

Basic information

• Original name: Immunosensing of the honeybee bacterial pathogen Melissococcus plutonius
• Authors: PASTUCHA, Matěj, Zdeněk FARKA, Veronika POLÁCHOVÁ, Zuzana MIKUŠOVÁ, Radka OBOŘILOVÁ and Petr SKLÁDAL.
• Edition: Joint Retreat 2019, 2019.

Other information

• Original language: English
• Type of outcome: Conference abstract
• Field of Study: 10406 Analytical chemistry
• Country of publisher: Czech Republic
• Confidentiality degree: is not subject to a state or trade secret
• WWW: URL
• Organization unit: Central European Institute of Technology
• ISBN: 978-80-210-9300-3
• Keywords in English: Antibody; Biosensor; Immunoassay; ELISA; Photon-upconversion nanoparticle; Melissococcus plutonius

Abstract

Western honeybee is an invaluable pollinator of many agricultural crops. European Foulbrood caused by the bacterium Melissococcus plutonius is one of the most serious honeybee diseases. In case of an outbreak, an effective method for the diagnosis is necessary, preferably sensitive enough to detect the pathogen before clinical symptoms manifest. In this work, we have exploited the antibody-based approach to develop new methods for the detection of the germ. No commercial antibodies were available; therefore, a new polyclonal antibody was prepared. This allowed us to develop an enzyme-linked immunosorbent assay (ELISA) for bacteria confirmation in laboratory with the limit of detection (LOD) of 7.1×104 CFU·mL−1. We further improved the sensitivity 400× by utilizing click-conjugated photon-upconversion nanoparticles (UCNPs) as a label for luminescence readout. In the next part of the research aiming for a portable detection system, we have developed an electrochemical immunosensor utilizing horseradish peroxidase as a label. With the LOD of 6.6×104 CFU·mL−1 and analysis time of 2 h, this allows sensitive in-field detection to prevent infection spreading and subsequent losses of honeybee colonies.

Links

• LQ1601, research and development project: Name: CEITEC 2020 (Acronym: CEITEC2020)

Investor: Ministry of Education, Youth and Sports of the CR

• TJ01000386, research and development project: Name: Imunostanovení pro diagnostiku hniloby včelího plodu

Investor: Technology Agency of the Czech Republic