Sensitive colorimetric immunoassay of Vibrio parahaemolyticus based on specific nonapeptide probe screening from a phage display library conjugated with MnO2 nanosheets with peroxidase-like activity

Pathogen detection continues to receive significant attention due to the harmful effects of pathogens on public health. Herein, specific nonapeptide-fusion proteins pVIII (pVIII fusion) were isolated from phage VQTVQIGSD (designated by the sequence of a fused foreign peptide), which was specifically...

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Bibliographic Details
Published in:Nanoscale 2018-02, Vol.10 (6), p.2825-2833
Main Authors: Liu, Pei, Han, Lei, Wang, Fei, Li, Xiangqian, Petrenko, Valery A, Liu, Aihua
Format: Article
Language:English
Subjects:
Antibodies
Bacteria
Colorimetry
Hydrogen peroxide
Immunoassay
Manganese dioxide
Nanosheets
Pathogens
Peroxidase
Phages
Proteins
Public health
Recognition
Seafood
Seawater
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Summary:Pathogen detection continues to receive significant attention due to the harmful effects of pathogens on public health. Herein, specific nonapeptide-fusion proteins pVIII (pVIII fusion) were isolated from phage VQTVQIGSD (designated by the sequence of a fused foreign peptide), which was specifically screened from the f8/9 landscape phage library against Vibrio parahaemolyticus (V. parahaemolyticus) in a high-throughput way. The as-prepared V. parahaemolyticus-specific recognition element is cheaper and more available than antibodies. Further, a highly sensitive colorimetric immunoassay for V. parahaemolyticus was established using pVIII fusion as capture probes coupled with protein-templated MnO2 nanosheets (NSs) as signal probes. In the presence of a target bacterium, V. parahaemolyticus, a sandwich-type complex of pVIII fusion–V. parahaemolyticus–MnO2 NS@pVIII fusion was formed through specific recognition of pVIII fusion and V. parahaemolyticus. The signal probes (MnO2 NSs) could catalyze the reaction of 3,3′,5,5′-tetramethylbenzidine and H2O2 to generate a colorimetric change. The proposed V. parahaemolyticus detection method demonstrated a wide detection range (20–104 colony-forming units (CFU) mL−1), low limit of detection (15 CFU mL−1), excellent selectivity, and high reliability for real marine samples, showing potential application in marine microbiological detection and control.
ISSN:2040-3364
2040-3372
DOI:10.1039/c7nr06633c