Determination of Aspergillus pathogens in agricultural products by a specific nanobody-polyclonal antibody sandwich ELISA
Abstract Aspergillus and its poisonous mycotoxins are distributed worldwide throughout the environment and are of particular interest in agriculture and food safety. In order to develop a specific method for rapid detection of Aspergillus flavus to forecast diseases and control aflatoxins, a nanobod...
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Nature Portfolio
2017
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oai:doaj.org-article:8f2e9df62fa047b3b2775e2bb62dcc3f2021-12-02T12:32:59ZDetermination of Aspergillus pathogens in agricultural products by a specific nanobody-polyclonal antibody sandwich ELISA10.1038/s41598-017-04195-62045-2322https://doaj.org/article/8f2e9df62fa047b3b2775e2bb62dcc3f2017-06-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-04195-6https://doaj.org/toc/2045-2322Abstract Aspergillus and its poisonous mycotoxins are distributed worldwide throughout the environment and are of particular interest in agriculture and food safety. In order to develop a specific method for rapid detection of Aspergillus flavus to forecast diseases and control aflatoxins, a nanobody, PO8-VHH, highly reactive to A. flavus was isolated from an immunized alpaca nanobody library by phage display. The nanobody was verified to bind to the components of extracellular and intracellular antigen from both A. flavus and A. parasiticus. To construct a sandwich format immunoassay, polyclonal antibodies against Aspergillus were raised with rabbits. Finally, a highly selective nanobody-polyclonal antibody sandwich enzyme-linked immunosorbent assay was optimized and developed. The results revealed that the detection limits of the two fungi were as low as 1 μg mL−1, and that it is able to detect fungal concentrations below to 2 μg mg−1 of peanut and maize grains in both artificially and naturally contaminated samples. Therefore, we here provided a rapid and simple method for monitoring Aspergillus spp. contamination in agricultural products.Ting WangPeiwu LiQi ZhangWen ZhangZhaowei ZhangTong WangTing HeNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-11 (2017) |
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Medicine R Science Q Ting Wang Peiwu Li Qi Zhang Wen Zhang Zhaowei Zhang Tong Wang Ting He Determination of Aspergillus pathogens in agricultural products by a specific nanobody-polyclonal antibody sandwich ELISA |
| description |
Abstract Aspergillus and its poisonous mycotoxins are distributed worldwide throughout the environment and are of particular interest in agriculture and food safety. In order to develop a specific method for rapid detection of Aspergillus flavus to forecast diseases and control aflatoxins, a nanobody, PO8-VHH, highly reactive to A. flavus was isolated from an immunized alpaca nanobody library by phage display. The nanobody was verified to bind to the components of extracellular and intracellular antigen from both A. flavus and A. parasiticus. To construct a sandwich format immunoassay, polyclonal antibodies against Aspergillus were raised with rabbits. Finally, a highly selective nanobody-polyclonal antibody sandwich enzyme-linked immunosorbent assay was optimized and developed. The results revealed that the detection limits of the two fungi were as low as 1 μg mL−1, and that it is able to detect fungal concentrations below to 2 μg mg−1 of peanut and maize grains in both artificially and naturally contaminated samples. Therefore, we here provided a rapid and simple method for monitoring Aspergillus spp. contamination in agricultural products. |
| format |
article |
| author |
Ting Wang Peiwu Li Qi Zhang Wen Zhang Zhaowei Zhang Tong Wang Ting He |
| author_facet |
Ting Wang Peiwu Li Qi Zhang Wen Zhang Zhaowei Zhang Tong Wang Ting He |
| author_sort |
Ting Wang |
| title |
Determination of Aspergillus pathogens in agricultural products by a specific nanobody-polyclonal antibody sandwich ELISA |
| title_short |
Determination of Aspergillus pathogens in agricultural products by a specific nanobody-polyclonal antibody sandwich ELISA |
| title_full |
Determination of Aspergillus pathogens in agricultural products by a specific nanobody-polyclonal antibody sandwich ELISA |
| title_fullStr |
Determination of Aspergillus pathogens in agricultural products by a specific nanobody-polyclonal antibody sandwich ELISA |
| title_full_unstemmed |
Determination of Aspergillus pathogens in agricultural products by a specific nanobody-polyclonal antibody sandwich ELISA |
| title_sort |
determination of aspergillus pathogens in agricultural products by a specific nanobody-polyclonal antibody sandwich elisa |
| publisher |
Nature Portfolio |
| publishDate |
2017 |
| url |
https://doaj.org/article/8f2e9df62fa047b3b2775e2bb62dcc3f |
| work_keys_str_mv |
AT tingwang determinationofaspergilluspathogensinagriculturalproductsbyaspecificnanobodypolyclonalantibodysandwichelisa AT peiwuli determinationofaspergilluspathogensinagriculturalproductsbyaspecificnanobodypolyclonalantibodysandwichelisa AT qizhang determinationofaspergilluspathogensinagriculturalproductsbyaspecificnanobodypolyclonalantibodysandwichelisa AT wenzhang determinationofaspergilluspathogensinagriculturalproductsbyaspecificnanobodypolyclonalantibodysandwichelisa AT zhaoweizhang determinationofaspergilluspathogensinagriculturalproductsbyaspecificnanobodypolyclonalantibodysandwichelisa AT tongwang determinationofaspergilluspathogensinagriculturalproductsbyaspecificnanobodypolyclonalantibodysandwichelisa AT tinghe determinationofaspergilluspathogensinagriculturalproductsbyaspecificnanobodypolyclonalantibodysandwichelisa |
| _version_ |
1718393932870582272 |