Amperometric Immunosensor for Rapid Detection of Honeybee
Pathogen Melissococcus Plutonius

J 2019

Amperometric Immunosensor for Rapid Detection of Honeybee Pathogen Melissococcus Plutonius

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

Základní údaje

Originální název

Amperometric Immunosensor for Rapid Detection of Honeybee Pathogen Melissococcus Plutonius

Autoři

MIKUŠOVÁ, Zuzana (703 Slovensko, domácí), Zdeněk FARKA (203 Česká republika, domácí), Matěj PASTUCHA (203 Česká republika, domácí), Veronika POLÁCHOVÁ (203 Česká republika, domácí), Radka OBOŘILOVÁ (203 Česká republika, domácí) a Petr SKLÁDAL (203 Česká republika, garant, domácí)

Vydání

Electroanalysis, WEINHEIM, WILEY-VCH Verlag GmbH, 2019, 1040-0397

Další údaje

Jazyk

angličtina

Typ výsledku

Článek v odborném periodiku

Obor

10406 Analytical chemistry

Stát vydavatele

Německo

Utajení

není předmětem státního či obchodního tajemství

Odkazy

URL

Impakt faktor

Impact factor: 2.544

Kód RIV

RIV/00216224:14740/19:00108372

Organizační jednotka

Středoevropský technologický institut

DOI

http://dx.doi.org/10.1002/elan.201900252

UT WoS

000474184300001

Klíčová slova anglicky

biosensor; amperometry; sandwich assay; antibody; European foulbrood; Apis mellifera

Štítky

CF NANO, rivok

Příznaky

Mezinárodní význam, Recenzováno

Změněno: 24. 10. 2024 11:50, Mgr. Adéla Pešková

Anotace

V originále

European foulbrood (EFB) is a honeybee larvae disease caused by a bacterium Melissococcus plutonius. An amperometric immunosensor based on a sandwich assay was developed for rapid point-of-care detection of this pathogen. An in-house made anti-Melissococcus antibody was immobilized to a gold surface of a screen-printed sensor via self-assembled monolayer of cysteamine activated with glutaraldehyde. The direct impedimetric detection of captured microbial cells was tested, however, a better performance was obtained after the formation of sandwich with the peroxidase-labeled antibody in the amperometric mode. The label-free assay was limited by higher non-specific binding. The limit of detection of the immunosensor was 6.6x10(4) CFU mL(-1) (colony-forming units) with wide linear range between 10(5) CFU mL(-1) and 10(9) CFU mL(-1). The whole analysis was completed within 2 h, which is shorter compared to common laboratory diagnostic tools, such as enzyme-linked immunosorbent assay or polymerase chain reaction. Furthermore, atomic force microscopy was used for confirmation of the bacteria presence on the electrode surface. The developed immunosensor was successfully employed in the analysis of real samples of honeybees and larvae. The achieved results demonstrate the potential of the amperometric immunosensor for practical in-field diagnosis of EFB, which can prevent infection spreading and connected losses of honeybee colonies.

Návaznosti

LM2015043, projekt VaV

Název: Česká infrastruktura pro integrativní strukturní biologii (Akronym: CIISB)

Investor: Ministerstvo školství, mládeže a tělovýchovy ČR, Czech Infrastructure for Integrative Structural Biology

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

MIKUŠOVÁ, Zuzana, Zdeněk FARKA, Matěj PASTUCHA, Veronika POLÁCHOVÁ, Radka OBOŘILOVÁ a Petr SKLÁDAL. Amperometric Immunosensor for Rapid Detection of Honeybee Pathogen Melissococcus Plutonius. Electroanalysis. WEINHEIM: WILEY-VCH Verlag GmbH, 2019, roč. 31, č. 10, s. 1969-1976. ISSN 1040-0397. Dostupné z: https://dx.doi.org/10.1002/elan.201900252.

@article{1573917,
   author = {Mikušová, Zuzana and Farka, Zdeněk and Pastucha, Matěj and Poláchová, Veronika and Obořilová, Radka and Skládal, Petr},
   article_location = {WEINHEIM},
   article_number = {10},
   doi = {http://dx.doi.org/10.1002/elan.201900252},
   keywords = {biosensor; amperometry; sandwich assay; antibody; European foulbrood; Apis mellifera},
   language = {eng},
   issn = {1040-0397},
   journal = {Electroanalysis},
   title = {Amperometric Immunosensor for Rapid Detection of Honeybee Pathogen Melissococcus Plutonius},
   url = {https://onlinelibrary.wiley.com/doi/abs/10.1002/elan.201900252},
   volume = {31},
   year = {2019}
}

TY  - JOUR
ID  - 1573917
AU  - Mikušová, Zuzana - Farka, Zdeněk - Pastucha, Matěj - Poláchová, Veronika - Obořilová, Radka - Skládal, Petr
PY  - 2019
TI  - Amperometric Immunosensor for Rapid Detection of Honeybee Pathogen Melissococcus Plutonius
JF  - Electroanalysis
VL  - 31
IS  - 10
SP  - 1969-1976
EP  - 1969-1976
PB  - WILEY-VCH Verlag GmbH
SN  - 10400397
KW  - biosensor
KW  - amperometry
KW  - sandwich assay
KW  - antibody
KW  - European foulbrood
KW  - Apis mellifera
UR  - https://onlinelibrary.wiley.com/doi/abs/10.1002/elan.201900252
L2  - https://onlinelibrary.wiley.com/doi/abs/10.1002/elan.201900252
N2  - European foulbrood (EFB) is a honeybee larvae disease caused by a bacterium Melissococcus plutonius. An amperometric immunosensor based on a sandwich assay was developed for rapid point-of-care detection of this pathogen. An in-house made anti-Melissococcus antibody was immobilized to a gold surface of a screen-printed sensor via self-assembled monolayer of cysteamine activated with glutaraldehyde. The direct impedimetric detection of captured microbial cells was tested, however, a better performance was obtained after the formation of sandwich with the peroxidase-labeled antibody in the amperometric mode. The label-free assay was limited by higher non-specific binding. The limit of detection of the immunosensor was 6.6x10(4) CFU mL(-1) (colony-forming units) with wide linear range between 10(5) CFU mL(-1) and 10(9) CFU mL(-1). The whole analysis was completed within 2 h, which is shorter compared to common laboratory diagnostic tools, such as enzyme-linked immunosorbent assay or polymerase chain reaction. Furthermore, atomic force microscopy was used for confirmation of the bacteria presence on the electrode surface. The developed immunosensor was successfully employed in the analysis of real samples of honeybees and larvae. The achieved results demonstrate the potential of the amperometric immunosensor for practical in-field diagnosis of EFB, which can prevent infection spreading and connected losses of honeybee colonies.
ER  -

MIKUŠOVÁ, Zuzana, Zdeněk FARKA, Matěj PASTUCHA, Veronika POLÁCHOVÁ, Radka OBOŘILOVÁ a Petr SKLÁDAL. Amperometric Immunosensor for Rapid Detection of Honeybee Pathogen Melissococcus Plutonius. \textit{Electroanalysis}. WEINHEIM: WILEY-VCH Verlag GmbH, 2019, roč.~31, č.~10, s.~1969-1976. ISSN~1040-0397. Dostupné z: https://dx.doi.org/10.1002/elan.201900252.

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