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Preparation of polyclonal antibody against a universal bacterial antigen OmpA deduced by bioinformatic analysis and preliminary evaluation of concentration effects on foodborne pathogens

Rapid and ultrasensitive microbial detection in actual samples have challenges because of target pathogen diversity and low abundance. In this study, we attempted to capture and concentrate multiple pathogens by combining magnetic beads with polyclonal antibodies against a universal antigen of ompA,...

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Autores principales: Wang, Lei, Ke, Yuehua, Li, Ye, Li, Yixuan, Yan, Yanfeng, Song, Yajun, Yang, Ruifu, Gao, Bo, Han, Yanping
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10208919/
https://www.ncbi.nlm.nih.gov/pubmed/37251856
http://dx.doi.org/10.1016/j.heliyon.2023.e16353
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author Wang, Lei
Ke, Yuehua
Li, Ye
Li, Yixuan
Yan, Yanfeng
Song, Yajun
Yang, Ruifu
Gao, Bo
Han, Yanping
author_facet Wang, Lei
Ke, Yuehua
Li, Ye
Li, Yixuan
Yan, Yanfeng
Song, Yajun
Yang, Ruifu
Gao, Bo
Han, Yanping
author_sort Wang, Lei
collection PubMed
description Rapid and ultrasensitive microbial detection in actual samples have challenges because of target pathogen diversity and low abundance. In this study, we attempted to capture and concentrate multiple pathogens by combining magnetic beads with polyclonal antibodies against a universal antigen of ompA, LAMOA-1, before further detection. A protein sequence consisting of 241 amino acids with spatial conformation similar to E. coli ompA was identified and expressed as a recombinant protein in prokaryotes according to the results of sequence alignment among 432 sequences of ompA belonging to intestinal bacteria from gram-negative bacteria. Purified from immunized rabbits, the anti-LAMOA-1 antibody was shown to effectively recognize 12 foodborne bacterial species. Antibody-conjugated beads were used to concentrate the bacteria when the bacterial concentration in artificially contaminated samples is between 10 and 100 CFU/mL, which shortens detection duration by 8–24 h. The enrichment strategy is potentially beneficial for detection of foodborne pathogens.
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spelling pubmed-102089192023-05-26 Preparation of polyclonal antibody against a universal bacterial antigen OmpA deduced by bioinformatic analysis and preliminary evaluation of concentration effects on foodborne pathogens Wang, Lei Ke, Yuehua Li, Ye Li, Yixuan Yan, Yanfeng Song, Yajun Yang, Ruifu Gao, Bo Han, Yanping Heliyon Research Article Rapid and ultrasensitive microbial detection in actual samples have challenges because of target pathogen diversity and low abundance. In this study, we attempted to capture and concentrate multiple pathogens by combining magnetic beads with polyclonal antibodies against a universal antigen of ompA, LAMOA-1, before further detection. A protein sequence consisting of 241 amino acids with spatial conformation similar to E. coli ompA was identified and expressed as a recombinant protein in prokaryotes according to the results of sequence alignment among 432 sequences of ompA belonging to intestinal bacteria from gram-negative bacteria. Purified from immunized rabbits, the anti-LAMOA-1 antibody was shown to effectively recognize 12 foodborne bacterial species. Antibody-conjugated beads were used to concentrate the bacteria when the bacterial concentration in artificially contaminated samples is between 10 and 100 CFU/mL, which shortens detection duration by 8–24 h. The enrichment strategy is potentially beneficial for detection of foodborne pathogens. Elsevier 2023-05-16 /pmc/articles/PMC10208919/ /pubmed/37251856 http://dx.doi.org/10.1016/j.heliyon.2023.e16353 Text en © 2023 The Authors https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Research Article
Wang, Lei
Ke, Yuehua
Li, Ye
Li, Yixuan
Yan, Yanfeng
Song, Yajun
Yang, Ruifu
Gao, Bo
Han, Yanping
Preparation of polyclonal antibody against a universal bacterial antigen OmpA deduced by bioinformatic analysis and preliminary evaluation of concentration effects on foodborne pathogens
title Preparation of polyclonal antibody against a universal bacterial antigen OmpA deduced by bioinformatic analysis and preliminary evaluation of concentration effects on foodborne pathogens
title_full Preparation of polyclonal antibody against a universal bacterial antigen OmpA deduced by bioinformatic analysis and preliminary evaluation of concentration effects on foodborne pathogens
title_fullStr Preparation of polyclonal antibody against a universal bacterial antigen OmpA deduced by bioinformatic analysis and preliminary evaluation of concentration effects on foodborne pathogens
title_full_unstemmed Preparation of polyclonal antibody against a universal bacterial antigen OmpA deduced by bioinformatic analysis and preliminary evaluation of concentration effects on foodborne pathogens
title_short Preparation of polyclonal antibody against a universal bacterial antigen OmpA deduced by bioinformatic analysis and preliminary evaluation of concentration effects on foodborne pathogens
title_sort preparation of polyclonal antibody against a universal bacterial antigen ompa deduced by bioinformatic analysis and preliminary evaluation of concentration effects on foodborne pathogens
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10208919/
https://www.ncbi.nlm.nih.gov/pubmed/37251856
http://dx.doi.org/10.1016/j.heliyon.2023.e16353
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