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@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.
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