DNA aptamer selection and detection of marine biotoxin 20 Methyl Spirolide G
| Autor: | Shivakumar R. Veerabhadraiah, Munna Singh Thakur, Praveena Bhatt, B. K. Bettadaiah, Srinivas Sistla, Monali Mukherjee |
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| Rok vydání: | 2021 |
| Předmět: |
Chromatography
20-methyl spirolide G Microscale thermophoresis Aptamer SELEX Aptamer Technique 010401 analytical chemistry 04 agricultural and veterinary sciences General Medicine Aptamers Nucleotide 040401 food science 01 natural sciences 0104 chemical sciences Analytical Chemistry chemistry.chemical_compound 0404 agricultural biotechnology chemistry Humans Marine Toxins Spiro Compounds Selection (genetic algorithm) Systematic evolution of ligands by exponential enrichment DNA Food Science |
| Zdroj: | Food Chemistry. 363:130332 |
| ISSN: | 0308-8146 |
| DOI: | 10.1016/j.foodchem.2021.130332 |
| Popis: | This study reports the selection of DNA aptamer for the detection of 20 Methyl Spirolide G (SPXG). After 10 rounds of selection, the enriched pool of aptamers specific to SPXG was cloned, sequenced and clustered into seven families based on similarity. Three sequences SPX1, SPX2 and SPX7, each belonging to different clades were further evaluated for their binding affinity. Surface plasmon resonance studies determined the highest affinity KD of 0.0345x10-8 M for aptamer SPX7. A label-free microscale thermophoresis-based aptasensing using SPX7 with highest affinity, indicated a linear detection range from 1.9 to 125000 pg/mL (LOD = 0.39 pg/mL; LOQ = 1.17 pg/mL). Spiking studies in simulated contaminated samples of mussel and scallop indicated recoveries in the range of 86 to 108%. Results of this study indicate the successful development of an aptamer for detection of SPXG at picogram levels. It also opens up avenues to develop other sensing platforms for detection of SPXG using the reported aptamer. |
| Databáze: | OpenAIRE |
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