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.

Basic information

Original name

Amperometric Immunosensor for Rapid Detection of Honeybee Pathogen Melissococcus Plutonius

Authors

MIKUŠOVÁ, Zuzana (703 Slovakia, belonging to the institution), Zdeněk FARKA (203 Czech Republic, belonging to the institution), Matěj PASTUCHA (203 Czech Republic, belonging to the institution), Veronika POLÁCHOVÁ (203 Czech Republic, belonging to the institution), Radka OBOŘILOVÁ (203 Czech Republic, belonging to the institution) and Petr SKLÁDAL (203 Czech Republic, guarantor, belonging to the institution)

Edition

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

Other information

Language

English

Type of outcome

Článek v odborném periodiku

Field of Study

10406 Analytical chemistry

Country of publisher

Germany

Confidentiality degree

není předmětem státního či obchodního tajemství

References:

URL

Impact factor

Impact factor: 2.544

RIV identification code

RIV/00216224:14740/19:00108372

Organization unit

Central European Institute of Technology

DOI

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

UT WoS

000474184300001

Keywords in English

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

Abstract

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.

Links

LM2015043, research and development project

Name: Česká infrastruktura pro integrativní strukturní biologii (Acronym: CIISB)

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

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

Citovat

MIKUŠOVÁ, Zuzana, Zdeněk FARKA, Matěj PASTUCHA, Veronika POLÁCHOVÁ, Radka OBOŘILOVÁ and Petr SKLÁDAL. Amperometric Immunosensor for Rapid Detection of Honeybee Pathogen Melissococcus Plutonius. Electroanalysis. WEINHEIM: WILEY-VCH Verlag GmbH, 2019, vol. 31, No 10, p. 1969-1976. ISSN 1040-0397. Available from: 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 and Petr SKLÁDAL. Amperometric Immunosensor for Rapid Detection of Honeybee Pathogen Melissococcus Plutonius. \textit{Electroanalysis}. WEINHEIM: WILEY-VCH Verlag GmbH, 2019, vol.~31, No~10, p.~1969-1976. ISSN~1040-0397. Available from: https://dx.doi.org/10.1002/elan.201900252.

Detailed Information on Publication Record