Amperometric Immunosensor for Rapid Detection of Honeybee Pathogen Melissococcus Plutonius

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00216224%3A14740%2F19%3A00108372" target="_blank" >RIV/00216224:14740/19:00108372 - isvavai.cz</a>

Výsledek na webu

<a href="https://onlinelibrary.wiley.com/doi/abs/10.1002/elan.201900252" target="_blank" >https://onlinelibrary.wiley.com/doi/abs/10.1002/elan.201900252</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1002/elan.201900252" target="_blank" >10.1002/elan.201900252</a>

Jazyk výsledku

angličtina

Název v původním jazyce

Amperometric Immunosensor for Rapid Detection of Honeybee Pathogen Melissococcus Plutonius

Popis výsledku v původním jazyce

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.

Název v anglickém jazyce

Amperometric Immunosensor for Rapid Detection of Honeybee Pathogen Melissococcus Plutonius

Popis výsledku anglicky

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.

Druh

J_imp - Článek v periodiku v databázi Web of Science

CEP obor

—

OECD FORD obor

10406 - Analytical chemistry

Projekt

Výsledek vznikl pri realizaci vícero projektů. Více informací v záložce Projekty.

Návaznosti

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)

Rok uplatnění

2019

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Název periodika

Electroanalysis

ISSN

1040-0397

e-ISSN

1521-4109

Svazek periodika

31

Číslo periodika v rámci svazku

10

Stát vydavatele periodika

DE - Spolková republika Německo

Počet stran výsledku

8

Strana od-do

1969-1976

Kód UT WoS článku

000474184300001

EID výsledku v databázi Scopus

2-s2.0-85068607764