One-pot electrochemical detection of foodborne pathogen based on in situ nucleic acid amplification and wash-free assay.
Mikrochim Acta
; 191(7): 431, 2024 06 29.
Article
en En
| MEDLINE
| ID: mdl-38951263
ABSTRACT
A signal amplification electrochemical biosensor chip was developed to integrate loop-mediated isothermal amplification (LAMP) based on in situ nucleic acid amplification and methyl blue (MB) serving as the hybridization redox indicator for sensitive and selective foodborne pathogen detection without a washing step. The electrochemical biosensor chip was designed by a screen-printed carbon electrode modified with gold nanoparticles (Au NPs) and covered with polydimethylsiloxane membrane to form a microcell. The primers of the target were immobilized on the Au NPs by covalent attachment for in situ amplification. The electroactive MB was used as the electrochemical signal reporter and embedded into the double-stranded DNA (dsDNA) amplicons generated by LAMP. Differential pulse voltammetry was introduced to survey the dsDNA hybridization with MB, which differentiates the specifically electrode-unbound and -bound labels without a washing step. Pyrene as the back-filling agent can further improve response signaling by reducing non-specific adsorption. This method is operationally simple, specific, and effective. The biosensor showed a detection linear range of 102-107 CFU mL-1 with the limit of detection of 17.7 CFU mL-1 within 40 min. This method showed promise for on-site testing of foodborne pathogens and could be integrated into an all-in-one device.
Palabras clave
Texto completo:
1
Colección:
01-internacional
Banco de datos:
MEDLINE
Asunto principal:
Técnicas Biosensibles
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Técnicas de Amplificación de Ácido Nucleico
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Nanopartículas del Metal
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Técnicas Electroquímicas
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Microbiología de Alimentos
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Oro
Idioma:
En
Revista:
Mikrochim Acta
/
Mikrochim. Acta (1966)
/
Mikrochimica acta (1966)
Año:
2024
Tipo del documento:
Article