Preparation of a Specific ssDNA Aptamer for Brevetoxin-2 Using SELEX
The existing assays for detecting brevetoxin (BTX) depend on expensive equipment with a professional operator or on an antibody with limited stability, which requires complex processes, a high cost, and a considerable amount of time. The development of an alternative detection probe is another promi...
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| Format: | Article |
| Language: | English |
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Wiley
2016-01-01
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| Series: | Journal of Analytical Methods in Chemistry |
| Online Access: | http://dx.doi.org/10.1155/2016/9241860 |
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| author | Rui-Yun Tian Chao Lin Shi-Yu Yu Sheng Gong Pan Hu Yan-Song Li Zong-Cheng Wu Yang Gao Yu Zhou Zeng-Shan Liu Hong-Lin Ren Shi-Ying Lu |
| author_facet | Rui-Yun Tian Chao Lin Shi-Yu Yu Sheng Gong Pan Hu Yan-Song Li Zong-Cheng Wu Yang Gao Yu Zhou Zeng-Shan Liu Hong-Lin Ren Shi-Ying Lu |
| author_sort | Rui-Yun Tian |
| collection | DOAJ |
| description | The existing assays for detecting brevetoxin (BTX) depend on expensive equipment with a professional operator or on an antibody with limited stability, which requires complex processes, a high cost, and a considerable amount of time. The development of an alternative detection probe is another promising research direction. This paper reports the use of aptamers binding to BTX-2 in an analytical assay using the systematic evolution of ligands by exponential enrichment (SELEX). After 12 rounds of selection, the secondary structures of 25 sequences were predicted. Compared to other aptamers, Bap5 has relatively high affinity with the lowest dissociation constant of 4.83 μM, and IC50 is 73.81 ng mL−1. A good linear regression formula of y=30.688x-7.329 with a coefficient correlation of R2 = 0.9798 was obtained using a biotin-avidin ELISA. Moreover, there is no cross-reaction with the detected marine toxins, except for BTX-2. Thus, Bap5 has potential to detect BTX-2 in shellfish in the future as a substitute for the recognition probe. |
| format | Article |
| id | doaj-art-7f4ba6bd03454d15a243af6ae3ecb913 |
| institution | Kabale University |
| issn | 2090-8865 2090-8873 |
| language | English |
| publishDate | 2016-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | Journal of Analytical Methods in Chemistry |
| spelling | doaj-art-7f4ba6bd03454d15a243af6ae3ecb9132025-02-03T05:51:21ZengWileyJournal of Analytical Methods in Chemistry2090-88652090-88732016-01-01201610.1155/2016/92418609241860Preparation of a Specific ssDNA Aptamer for Brevetoxin-2 Using SELEXRui-Yun Tian0Chao Lin1Shi-Yu Yu2Sheng Gong3Pan Hu4Yan-Song Li5Zong-Cheng Wu6Yang Gao7Yu Zhou8Zeng-Shan Liu9Hong-Lin Ren10Shi-Ying Lu11Key Laboratory of Zoonosis Research, Ministry of Education, Institute of Zoonosis, College of Veterinary Medicine, Jilin University, Changchun 130062, ChinaKey Laboratory of Zoonosis Research, Ministry of Education, Institute of Zoonosis, College of Veterinary Medicine, Jilin University, Changchun 130062, ChinaFuqing Entry-Exit Inspection and Quarantine Bureau, Port District, Qingrong Road, Fuqing, Fujian 350300, ChinaKey Laboratory of Zoonosis Research, Ministry of Education, Institute of Zoonosis, College of Veterinary Medicine, Jilin University, Changchun 130062, ChinaKey Laboratory of Zoonosis Research, Ministry of Education, Institute of Zoonosis, College of Veterinary Medicine, Jilin University, Changchun 130062, ChinaKey Laboratory of Zoonosis Research, Ministry of Education, Institute of Zoonosis, College of Veterinary Medicine, Jilin University, Changchun 130062, ChinaKey Laboratory of Zoonosis Research, Ministry of Education, Institute of Zoonosis, College of Veterinary Medicine, Jilin University, Changchun 130062, ChinaKey Laboratory of Zoonosis Research, Ministry of Education, Institute of Zoonosis, College of Veterinary Medicine, Jilin University, Changchun 130062, ChinaKey Laboratory of Zoonosis Research, Ministry of Education, Institute of Zoonosis, College of Veterinary Medicine, Jilin University, Changchun 130062, ChinaKey Laboratory of Zoonosis Research, Ministry of Education, Institute of Zoonosis, College of Veterinary Medicine, Jilin University, Changchun 130062, ChinaKey Laboratory of Zoonosis Research, Ministry of Education, Institute of Zoonosis, College of Veterinary Medicine, Jilin University, Changchun 130062, ChinaKey Laboratory of Zoonosis Research, Ministry of Education, Institute of Zoonosis, College of Veterinary Medicine, Jilin University, Changchun 130062, ChinaThe existing assays for detecting brevetoxin (BTX) depend on expensive equipment with a professional operator or on an antibody with limited stability, which requires complex processes, a high cost, and a considerable amount of time. The development of an alternative detection probe is another promising research direction. This paper reports the use of aptamers binding to BTX-2 in an analytical assay using the systematic evolution of ligands by exponential enrichment (SELEX). After 12 rounds of selection, the secondary structures of 25 sequences were predicted. Compared to other aptamers, Bap5 has relatively high affinity with the lowest dissociation constant of 4.83 μM, and IC50 is 73.81 ng mL−1. A good linear regression formula of y=30.688x-7.329 with a coefficient correlation of R2 = 0.9798 was obtained using a biotin-avidin ELISA. Moreover, there is no cross-reaction with the detected marine toxins, except for BTX-2. Thus, Bap5 has potential to detect BTX-2 in shellfish in the future as a substitute for the recognition probe.http://dx.doi.org/10.1155/2016/9241860 |
| spellingShingle | Rui-Yun Tian Chao Lin Shi-Yu Yu Sheng Gong Pan Hu Yan-Song Li Zong-Cheng Wu Yang Gao Yu Zhou Zeng-Shan Liu Hong-Lin Ren Shi-Ying Lu Preparation of a Specific ssDNA Aptamer for Brevetoxin-2 Using SELEX Journal of Analytical Methods in Chemistry |
| title | Preparation of a Specific ssDNA Aptamer for Brevetoxin-2 Using SELEX |
| title_full | Preparation of a Specific ssDNA Aptamer for Brevetoxin-2 Using SELEX |
| title_fullStr | Preparation of a Specific ssDNA Aptamer for Brevetoxin-2 Using SELEX |
| title_full_unstemmed | Preparation of a Specific ssDNA Aptamer for Brevetoxin-2 Using SELEX |
| title_short | Preparation of a Specific ssDNA Aptamer for Brevetoxin-2 Using SELEX |
| title_sort | preparation of a specific ssdna aptamer for brevetoxin 2 using selex |
| url | http://dx.doi.org/10.1155/2016/9241860 |
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