Nanobody-based immunosensor for the detection of H. pylori in saliva

Helicobacter pylori (H. pylori) infection is highly prevalent worldwide, affecting more than 43% of world population. The infection can be transmitted through different routes, like oral-oral, fecal-oral, and gastric-oral. Electrochemical sensors play a crucial role in the early detection of various...

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Published in:Biosensors & bioelectronics 2024-09, Vol.260, p.116423-116423, Article 116423
Main Authors: Ahmad, Mohammad IA, Amorim, Célia G., Abu Qatouseh, Luay F., Montenegro, Maria C.B.S.M.
Format: Article
Language:English
Subjects:
Antibodies, Immobilized - chemistry
Antibodies, Immobilized - immunology
Antigens, Bacterial - analysis
Antigens, Bacterial - immunology
Antigens, Bacterial - isolation & purification
Biosensing Techniques - instrumentation
Biosensors
Diagnostic techniques and procedures
Electrochemical immunoassay
Electrochemical Techniques
Gold - chemistry
H. pylori
Helicobacter Infections - diagnosis
Helicobacter Infections - immunology
Helicobacter Infections - microbiology
Helicobacter pylori - immunology
Helicobacter pylori - isolation & purification
Humans
Immunoassay - methods
Limit of Detection
Nanobodies
Saliva - chemistry
Saliva - microbiology
Single-Domain Antibodies - chemistry
Single-Domain Antibodies - immunology
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Summary:Helicobacter pylori (H. pylori) infection is highly prevalent worldwide, affecting more than 43% of world population. The infection can be transmitted through different routes, like oral-oral, fecal-oral, and gastric-oral. Electrochemical sensors play a crucial role in the early detection of various substances, including biomolecules. In this study, the development of nanobody (Nb)-based immunosensor for the detection of H. pylori antigens in saliva samples was investigated. The D2_Nb was isolated and characterized using Western blot and ELISA and employed in the fabrication of the immunosensor. The sensor was prepared using gold screen-printed electrodes, with the immobilization of Nb achieved through chemical linkage using cysteamine-glutaraldehyde. The surface of the electrode was characterized using EIS, FTIR and SEM. Initially, the Nb-based immunosensor's performance was evaluated through cyclic voltammetry (CV), differential pulse voltammetry (DPV), and square wave voltammetry (SWV). The sensor exhibited excellent linearity with an R2 value of 0.96. However, further assessment with the DPV technique revealed both a low limit of detection (5.9 ng/mL,
ISSN:0956-5663
1873-4235
DOI:10.1016/j.bios.2024.116423