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目的:建立一种快速、灵敏、特异的基于适配子及Dot-ELISA的氯霉素残留检测方法。方法:选取一种能与氯霉素特异性结合的DNA适配分子,人工合成并用生物素标记。把含有不同浓度氯霉素的样品结合于硝酸纤维膜上,然后与生物素标记的适配子于55℃孵育1 h,洗脱,接着与亲和素标记的辣根过氧化物酶孵育,DAB显色。结果:成功建立了基于适配分子的Dot-ELISA检测氯霉素残留的方法,可以快速检测牛奶等食品中的氯霉素残留,检测灵敏度达10-7g/L。结论:所建立的基于适配分子的Dot-ELISA方法可快速检测氯霉素残留,且具有灵敏性高、特异性强等特点,可作为抗生素残留筛查的较好选择,有广阔的应用前景。
OBJECTIVE: To establish a rapid, sensitive and specific method for detection of chloramphenicol residues based on aptamers and Dot-ELISA. METHODS: A DNA adapter molecule that specifically binds to chloramphenicol was synthesized and labeled with biotin. Samples containing different concentrations of chloramphenicol were bound to nitrocellulose and then incubated with biotinylated aptamers for 1 h at 55 ° C, followed by incubation with avidin-labeled horseradish peroxidase (HRP) DAB color. Results: Dot-ELISA based on adapter molecules was successfully established to detect chloramphenicol residues. The method can detect chloramphenicol residues in milk and other foods rapidly with the detection sensitivity of 10-7g / L. Conclusion: Dot-ELISA based on adapter molecule can detect chloramphenicol residues quickly and has the characteristics of high sensitivity and specificity. It can be used as a good choice of antibiotic residue screening and has broad application prospects .