Immunosensing of the honeybee bacterial pathogen Melissococcus
plutonius

a 2019

Immunosensing of the honeybee bacterial pathogen Melissococcus plutonius

PASTUCHA, Matěj, Zdeněk FARKA, Veronika POLÁCHOVÁ, Zuzana MIKUŠOVÁ, Radka OBOŘILOVÁ et. al.

Základní údaje

Originální název

Immunosensing of the honeybee bacterial pathogen Melissococcus plutonius

Autoři

PASTUCHA, Matěj, Zdeněk FARKA, Veronika POLÁCHOVÁ, Zuzana MIKUŠOVÁ, Radka OBOŘILOVÁ a Petr SKLÁDAL

Vydání

Joint Retreat 2019, 2019

Další údaje

Jazyk

angličtina

Typ výsledku

Konferenční abstrakt

Obor

10406 Analytical chemistry

Stát vydavatele

Česká republika

Utajení

není předmětem státního či obchodního tajemství

Odkazy

URL

Organizační jednotka

Středoevropský technologický institut

ISBN

978-80-210-9300-3

Klíčová slova anglicky

Antibody; Biosensor; Immunoassay; ELISA; Photon-upconversion nanoparticle; Melissococcus plutonius

Změněno: 5. 7. 2019 10:41, doc. Mgr. Zdeněk Farka, Ph.D.

Anotace

V originále

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.

Návaznosti

LQ1601, projekt VaV

Název: CEITEC 2020 (Akronym: CEITEC2020)

Investor: Ministerstvo školství, mládeže a tělovýchovy ČR, CEITEC 2020

TJ01000386, projekt VaV

Název: Imunostanovení pro diagnostiku hniloby včelího plodu

Investor: Technologická agentura ČR, Imunostanovení pro diagnostiku hniloby včelího plodu

Citovat

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

@proceedings{1544338,
   author = {Pastucha, Matěj and Farka, Zdeněk and Poláchová, Veronika and Mikušová, Zuzana and Obořilová, Radka and Skládal, Petr},
   booktitle = {Joint Retreat 2019},
   keywords = {Antibody; Biosensor; Immunoassay; ELISA; Photon-upconversion nanoparticle; Melissococcus plutonius},
   language = {eng},
   isbn = {978-80-210-9300-3},
   title = {Immunosensing of the honeybee bacterial pathogen Melissococcus plutonius},
   url = {http://phdretreat.ceitec.cz/download/220},
   year = {2019}
}

TY  - CONF
ID  - 1544338
AU  - Pastucha, Matěj - Farka, Zdeněk - Poláchová, Veronika - Mikušová, Zuzana - Obořilová, Radka - Skládal, Petr
PY  - 2019
TI  - Immunosensing of the honeybee bacterial pathogen Melissococcus plutonius
SN  - 9788021093003
KW  - Antibody
KW  - Biosensor
KW  - Immunoassay
KW  - ELISA
KW  - Photon-upconversion nanoparticle
KW  - Melissococcus plutonius
UR  - http://phdretreat.ceitec.cz/download/220
N2  - 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.
ER  -

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

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