One-pot electrochemical detection of foodborne pathogen based on in situ nucleic acid amplification and wash-free assay.


Journal

Mikrochimica acta
ISSN: 1436-5073
Titre abrégé: Mikrochim Acta
Pays: Austria
ID NLM: 7808782

Informations de publication

Date de publication:
29 Jun 2024
Historique:
received: 03 04 2024
accepted: 10 06 2024
medline: 2 7 2024
pubmed: 2 7 2024
entrez: 1 7 2024
Statut: epublish

Résumé

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 10

Identifiants

pubmed: 38951263
doi: 10.1007/s00604-024-06500-3
pii: 10.1007/s00604-024-06500-3
doi:

Substances chimiques

Gold 7440-57-5
DNA, Bacterial 0

Types de publication

Journal Article

Langues

eng

Sous-ensembles de citation

IM

Pagination

431

Subventions

Organisme : Science and Technology Benefiting the People Demonstration Project of Qingdao
ID : 22-2-7-smjk-2-nsh
Organisme : Key Project of Shandong Province Natural Science Foundation
ID : ZR2020KH030

Informations de copyright

© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Austria, part of Springer Nature.

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Auteurs

Hao Gong (H)

Qingdao Nucleic Acid Rapid Testing International Science and Technology Cooperation Base, College of Life SciencesDepartment of Pathogenic Biology, School of Basic MedicineDepartment of Clinical Laboratory, the Affiliated Hospital of Qingdao University, Qingdao University, Qingdao, 266071, People's Republic of China.

Haoran Wang (H)

Shandong Provincial Key Laboratory of Biochemical Engineering, Qingdao Key Laboratory of Nucleic Acid Rapid Detection, Sino-UAE International Cooperative Joint Laboratory of Pathogenic Microorganism Rapid Detection, College of Biological Engineering, Qingdao Nucleic Acid Rapid Detection Engineering Research Center, Qingdao University of Science and Technology, Qingdao, 266042, People's Republic of China.

Yanli Ji (Y)

Qingdao JianMa Gene Technology Co., Ltd, Qingdao, 266114, People's Republic of China.

Longqiang Xu (L)

Department of Clinical Laboratory, The Affiliated Hospital of Qingdao University, Qingdao, 266071, People's Republic of China. xulongqiang75@sina.com.

Cuiping Ma (C)

Shandong Provincial Key Laboratory of Biochemical Engineering, Qingdao Key Laboratory of Nucleic Acid Rapid Detection, Sino-UAE International Cooperative Joint Laboratory of Pathogenic Microorganism Rapid Detection, College of Biological Engineering, Qingdao Nucleic Acid Rapid Detection Engineering Research Center, Qingdao University of Science and Technology, Qingdao, 266042, People's Republic of China.

Chao Shi (C)

Qingdao Nucleic Acid Rapid Testing International Science and Technology Cooperation Base, College of Life SciencesDepartment of Pathogenic Biology, School of Basic MedicineDepartment of Clinical Laboratory, the Affiliated Hospital of Qingdao University, Qingdao University, Qingdao, 266071, People's Republic of China. sc169@163.com.
Qingdao JianMa Gene Technology Co., Ltd, Qingdao, 266114, People's Republic of China. sc169@163.com.

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