Screening of Single-Stranded DNA Aptamer Specific for Florfenicol and Application in Detection of Food Safety
In this work, the single-stranded DNA (ssDNA) aptamers specific to florfenicol (FF) and having a high binding affinity were prepared using the magnetic bead-based systematic evolution of ligands by the exponential enrichment technique (MB-SELEX). After 10 rounds of the MB-SELEX screening, aptamers t...
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| Format: | Article |
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MDPI AG
2022-09-01
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| Series: | Biosensors |
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| Online Access: | https://www.mdpi.com/2079-6374/12/9/701 |
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| author | Minghui Shi Ruobing Liu Fuyuan Zhang Bimal Chitrakar Xianghong Wang |
| author_facet | Minghui Shi Ruobing Liu Fuyuan Zhang Bimal Chitrakar Xianghong Wang |
| author_sort | Minghui Shi |
| collection | DOAJ |
| description | In this work, the single-stranded DNA (ssDNA) aptamers specific to florfenicol (FF) and having a high binding affinity were prepared using the magnetic bead-based systematic evolution of ligands by the exponential enrichment technique (MB-SELEX). After 10 rounds of the MB-SELEX screening, aptamers that can simultaneously recognize FF and its metabolite florfenicol amine (FFA) were obtained. The aptamer with the lowest dissociation constant (K<sub>d</sub>) was truncated and optimized based on a secondary structure analysis. The optimal aptamer selected was Apt-14t, with a length of 43 nt, and its dissociation constant was 4.66 ± 0.75 nM, which was about 7 times higher than that of the full-length sequence. The potential binding sites and interactions with FF were demonstrated by molecular docking simulations. In addition, a colorimetric strategy for nanogold aptamers was constructed. The linear detection range of this method was 0.00128–500 ng/mL and the actual detection limit was 0.00128 ng/mL. Using this strategy to detect florfenicol in actual milk and eggs samples, the spiked recoveries were 88.9–123.1% and 84.0–112.2%, respectively, and the relative standard deviation was less than 5.6%, showing high accuracy. |
| first_indexed | 2024-03-10T00:35:17Z |
| format | Article |
| id | doaj.art-53eefd1a2a64402ca737474980890956 |
| institution | Directory Open Access Journal |
| issn | 2079-6374 |
| language | English |
| last_indexed | 2024-03-10T00:35:17Z |
| publishDate | 2022-09-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Biosensors |
| spelling | doaj.art-53eefd1a2a64402ca7374749808909562023-11-23T15:17:42ZengMDPI AGBiosensors2079-63742022-09-0112970110.3390/bios12090701Screening of Single-Stranded DNA Aptamer Specific for Florfenicol and Application in Detection of Food SafetyMinghui Shi0Ruobing Liu1Fuyuan Zhang2Bimal Chitrakar3Xianghong Wang4College of Food Science and Technology, Hebei Agricultural University, Baoding 071001, ChinaCollege of Food Science and Technology, Hebei Agricultural University, Baoding 071001, ChinaCollege of Food Science and Technology, Hebei Agricultural University, Baoding 071001, ChinaCollege of Food Science and Technology, Hebei Agricultural University, Baoding 071001, ChinaCollege of Food Science and Technology, Hebei Agricultural University, Baoding 071001, ChinaIn this work, the single-stranded DNA (ssDNA) aptamers specific to florfenicol (FF) and having a high binding affinity were prepared using the magnetic bead-based systematic evolution of ligands by the exponential enrichment technique (MB-SELEX). After 10 rounds of the MB-SELEX screening, aptamers that can simultaneously recognize FF and its metabolite florfenicol amine (FFA) were obtained. The aptamer with the lowest dissociation constant (K<sub>d</sub>) was truncated and optimized based on a secondary structure analysis. The optimal aptamer selected was Apt-14t, with a length of 43 nt, and its dissociation constant was 4.66 ± 0.75 nM, which was about 7 times higher than that of the full-length sequence. The potential binding sites and interactions with FF were demonstrated by molecular docking simulations. In addition, a colorimetric strategy for nanogold aptamers was constructed. The linear detection range of this method was 0.00128–500 ng/mL and the actual detection limit was 0.00128 ng/mL. Using this strategy to detect florfenicol in actual milk and eggs samples, the spiked recoveries were 88.9–123.1% and 84.0–112.2%, respectively, and the relative standard deviation was less than 5.6%, showing high accuracy.https://www.mdpi.com/2079-6374/12/9/701florfenicolSELEXaptamerAuNPsmolecular docking |
| spellingShingle | Minghui Shi Ruobing Liu Fuyuan Zhang Bimal Chitrakar Xianghong Wang Screening of Single-Stranded DNA Aptamer Specific for Florfenicol and Application in Detection of Food Safety Biosensors florfenicol SELEX aptamer AuNPs molecular docking |
| title | Screening of Single-Stranded DNA Aptamer Specific for Florfenicol and Application in Detection of Food Safety |
| title_full | Screening of Single-Stranded DNA Aptamer Specific for Florfenicol and Application in Detection of Food Safety |
| title_fullStr | Screening of Single-Stranded DNA Aptamer Specific for Florfenicol and Application in Detection of Food Safety |
| title_full_unstemmed | Screening of Single-Stranded DNA Aptamer Specific for Florfenicol and Application in Detection of Food Safety |
| title_short | Screening of Single-Stranded DNA Aptamer Specific for Florfenicol and Application in Detection of Food Safety |
| title_sort | screening of single stranded dna aptamer specific for florfenicol and application in detection of food safety |
| topic | florfenicol SELEX aptamer AuNPs molecular docking |
| url | https://www.mdpi.com/2079-6374/12/9/701 |
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