Application of colloidal gold immunoassay strips for the rapid detection of potato Y virus
Potato Y virus (PVY) is a bacterial virus that seriously jeopardizes the growth of tobacco. In order to achieve rapid detection of PVY, monoclonal antibodies to PVY-CP protein were prepared and characterized, and colloidal gold immunochromatographic test strips that can be used to detect PVY were es...
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| Format: | Article |
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EDP Sciences
2024-01-01
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| Series: | BIO Web of Conferences |
| Online Access: | https://www.bio-conferences.org/articles/bioconf/pdf/2024/61/bioconf_isaeb2024_03009.pdf |
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| author | Wang Chunqiong Dan Chen Zhang Xiaowei Zhu Dan Cai Jieyun Long Jie Zhang Ke Meng Hongming Haowei Sun Kai Liu Zeng Yanbo |
| author_facet | Wang Chunqiong Dan Chen Zhang Xiaowei Zhu Dan Cai Jieyun Long Jie Zhang Ke Meng Hongming Haowei Sun Kai Liu Zeng Yanbo |
| author_sort | Wang Chunqiong |
| collection | DOAJ |
| description | Potato Y virus (PVY) is a bacterial virus that seriously jeopardizes the growth of tobacco. In order to achieve rapid detection of PVY, monoclonal antibodies to PVY-CP protein were prepared and characterized, and colloidal gold immunochromatographic test strips that can be used to detect PVY were established. In this study, we constructed the PVY-CP protein expression plasmid pET28a-PVY-CP, transformed it into Escherichia coli BL21 (DE3) receptor cells to induce the expression of the target protein, and then further purified it as an immunogen to screen hybridoma cells that can stably secrete monoclonal antibody against PVY through cell fusion with hybridoma cells, prepared ascites, and purified the monoclonal antibody by using the caprylic acid saturated ammonium sulfate method. The monoclonal antibody was purified using the method of ammonium octanoate saturated sulfate, labeled with colloidal gold, and the colloidal gold immunochromatographic test strip was established by optimizing the reaction conditions, and the detectability, accuracy and specificity of the test strip were evaluated. The detection limit of the test strip for PVY-CP protein was 1 μg/mL, and there was no cross-reactivity with tobacco bunchy top virus, tomato spotted wilt virus, and tobacco mosaic virus. Comparison of the prepared colloidal gold test strips and the RT-PCR method for actual samples showed that the total conformity rate of the two was 86.67%, and the positive conformity rate was 90%. |
| format | Article |
| id | doaj-art-cd6af50126544319a4b31bd44aed7614 |
| institution | OA Journals |
| issn | 2117-4458 |
| language | English |
| publishDate | 2024-01-01 |
| publisher | EDP Sciences |
| record_format | Article |
| series | BIO Web of Conferences |
| spelling | doaj-art-cd6af50126544319a4b31bd44aed76142025-08-20T02:36:09ZengEDP SciencesBIO Web of Conferences2117-44582024-01-011420300910.1051/bioconf/202414203009bioconf_isaeb2024_03009Application of colloidal gold immunoassay strips for the rapid detection of potato Y virusWang Chunqiong0Dan Chen1Zhang Xiaowei2Zhu Dan3Cai Jieyun4Long Jie5Zhang Ke6Meng Hongming7Haowei Sun8Kai Liu9Zeng Yanbo10Yunnan Tobacco Quality Supervision and Inspection StationYunnan Tobacco Quality Supervision and Inspection StationYunnan Tobacco Quality Supervision and Inspection StationKunming Cigarette Factory of Hongyun Honghe GroupYunnan Tobacco Quality Supervision and Inspection StationYunnan Tobacco Quality Supervision and Inspection StationYunnan Tobacco Quality Supervision and Inspection StationYunnan Tobacco Quality Supervision and Inspection StationYunnan Tobacco Quality Supervision and Inspection StationYunnan Tobacco Quality Supervision and Inspection StationYunnan University for Nationalities, KunmingPotato Y virus (PVY) is a bacterial virus that seriously jeopardizes the growth of tobacco. In order to achieve rapid detection of PVY, monoclonal antibodies to PVY-CP protein were prepared and characterized, and colloidal gold immunochromatographic test strips that can be used to detect PVY were established. In this study, we constructed the PVY-CP protein expression plasmid pET28a-PVY-CP, transformed it into Escherichia coli BL21 (DE3) receptor cells to induce the expression of the target protein, and then further purified it as an immunogen to screen hybridoma cells that can stably secrete monoclonal antibody against PVY through cell fusion with hybridoma cells, prepared ascites, and purified the monoclonal antibody by using the caprylic acid saturated ammonium sulfate method. The monoclonal antibody was purified using the method of ammonium octanoate saturated sulfate, labeled with colloidal gold, and the colloidal gold immunochromatographic test strip was established by optimizing the reaction conditions, and the detectability, accuracy and specificity of the test strip were evaluated. The detection limit of the test strip for PVY-CP protein was 1 μg/mL, and there was no cross-reactivity with tobacco bunchy top virus, tomato spotted wilt virus, and tobacco mosaic virus. Comparison of the prepared colloidal gold test strips and the RT-PCR method for actual samples showed that the total conformity rate of the two was 86.67%, and the positive conformity rate was 90%.https://www.bio-conferences.org/articles/bioconf/pdf/2024/61/bioconf_isaeb2024_03009.pdf |
| spellingShingle | Wang Chunqiong Dan Chen Zhang Xiaowei Zhu Dan Cai Jieyun Long Jie Zhang Ke Meng Hongming Haowei Sun Kai Liu Zeng Yanbo Application of colloidal gold immunoassay strips for the rapid detection of potato Y virus BIO Web of Conferences |
| title | Application of colloidal gold immunoassay strips for the rapid detection of potato Y virus |
| title_full | Application of colloidal gold immunoassay strips for the rapid detection of potato Y virus |
| title_fullStr | Application of colloidal gold immunoassay strips for the rapid detection of potato Y virus |
| title_full_unstemmed | Application of colloidal gold immunoassay strips for the rapid detection of potato Y virus |
| title_short | Application of colloidal gold immunoassay strips for the rapid detection of potato Y virus |
| title_sort | application of colloidal gold immunoassay strips for the rapid detection of potato y virus |
| url | https://www.bio-conferences.org/articles/bioconf/pdf/2024/61/bioconf_isaeb2024_03009.pdf |
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