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Development of a double-antibody sandwich ELISA for rapid detection of Bacillus Cereus in food
Bacillus cereus is increasingly recognized as one of the major causes of food poisoning in the industrialized world. In this paper, we describe a sensitive double-antibody sandwich enzyme-linked immunosorbent assay (ELISA) that was developed for rapid detection of B. cereus in food to minimize the r...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group
2016
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4792147/ https://www.ncbi.nlm.nih.gov/pubmed/26976753 http://dx.doi.org/10.1038/srep16092 |
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author | Zhu, Longjiao He, Jing Cao, Xiaohan Huang, Kunlun Luo, Yunbo Xu, Wentao |
author_facet | Zhu, Longjiao He, Jing Cao, Xiaohan Huang, Kunlun Luo, Yunbo Xu, Wentao |
author_sort | Zhu, Longjiao |
collection | PubMed |
description | Bacillus cereus is increasingly recognized as one of the major causes of food poisoning in the industrialized world. In this paper, we describe a sensitive double-antibody sandwich enzyme-linked immunosorbent assay (ELISA) that was developed for rapid detection of B. cereus in food to minimize the risk of contamination. The polyclonal antibody (pAb) and monoclonal antibodies (mAbs) specific to B. cereus were generated from rabbit antiserum and mouse ascites, respectively, using the octanoic acid/saturated ammonium sulfate precipitation method and protein A-sepharose columns. IgG-isotype mAbs were specially developed to undergo a novel peripheral multiple sites immunization for rapid gain of hybridomas and a subtractive screen was used to eliminate cross reactivity with closely related species such as Bacillus thuringiensis, B. subtilis, B. licheniformis and B. perfringens. The linear detection range of the method was approximately 1 × 10(4)–2.8 × 10(6) cells/mL with a detection limit (LOD) of 0.9 × 10(3) cells/mL. The assay was able to detect B. cereus when the samples were prepared in meat with various pathogens. The newly developed analytical method provides a rapid method to sensitively detect B. cereus in food specimens. |
format | Online Article Text |
id | pubmed-4792147 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | Nature Publishing Group |
record_format | MEDLINE/PubMed |
spelling | pubmed-47921472016-03-16 Development of a double-antibody sandwich ELISA for rapid detection of Bacillus Cereus in food Zhu, Longjiao He, Jing Cao, Xiaohan Huang, Kunlun Luo, Yunbo Xu, Wentao Sci Rep Article Bacillus cereus is increasingly recognized as one of the major causes of food poisoning in the industrialized world. In this paper, we describe a sensitive double-antibody sandwich enzyme-linked immunosorbent assay (ELISA) that was developed for rapid detection of B. cereus in food to minimize the risk of contamination. The polyclonal antibody (pAb) and monoclonal antibodies (mAbs) specific to B. cereus were generated from rabbit antiserum and mouse ascites, respectively, using the octanoic acid/saturated ammonium sulfate precipitation method and protein A-sepharose columns. IgG-isotype mAbs were specially developed to undergo a novel peripheral multiple sites immunization for rapid gain of hybridomas and a subtractive screen was used to eliminate cross reactivity with closely related species such as Bacillus thuringiensis, B. subtilis, B. licheniformis and B. perfringens. The linear detection range of the method was approximately 1 × 10(4)–2.8 × 10(6) cells/mL with a detection limit (LOD) of 0.9 × 10(3) cells/mL. The assay was able to detect B. cereus when the samples were prepared in meat with various pathogens. The newly developed analytical method provides a rapid method to sensitively detect B. cereus in food specimens. Nature Publishing Group 2016-03-15 /pmc/articles/PMC4792147/ /pubmed/26976753 http://dx.doi.org/10.1038/srep16092 Text en Copyright © 2015, Macmillan Publishers Limited http://creativecommons.org/licenses/by/4.0/ This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/ |
spellingShingle | Article Zhu, Longjiao He, Jing Cao, Xiaohan Huang, Kunlun Luo, Yunbo Xu, Wentao Development of a double-antibody sandwich ELISA for rapid detection of Bacillus Cereus in food |
title | Development of a double-antibody sandwich ELISA for rapid detection of Bacillus Cereus in food |
title_full | Development of a double-antibody sandwich ELISA for rapid detection of Bacillus Cereus in food |
title_fullStr | Development of a double-antibody sandwich ELISA for rapid detection of Bacillus Cereus in food |
title_full_unstemmed | Development of a double-antibody sandwich ELISA for rapid detection of Bacillus Cereus in food |
title_short | Development of a double-antibody sandwich ELISA for rapid detection of Bacillus Cereus in food |
title_sort | development of a double-antibody sandwich elisa for rapid detection of bacillus cereus in food |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4792147/ https://www.ncbi.nlm.nih.gov/pubmed/26976753 http://dx.doi.org/10.1038/srep16092 |
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