Yang Qingbao, Qi Yanhua, Zhou Jingming, Chen Yumei, Liang Chao, Liu Zhanxiang, Zhang Xiaoli, Wang Aiping
School of Life Sciences, Zhengzhou University Zhengzhou 450001 Henan China
RSC Adv. 2021 Jun 22;11(36):22005-22013. doi: 10.1039/d1ra03065e. eCollection 2021 Jun 21.
Fleroxacin (FLE) is a broad-spectrum fluoroquinolone antibiotic widely used in animal husbandry, veterinary medicine and aquaculture. Eating animal-derived foods with FLE residues can cause allergies, poisoning or drug resistance. The water-soluble QDs (CdSe/ZnS) and anti-FLE monoclonal antibody (mAb) were used to prepare a fluorescent probe by the method of -(3-dimethylaminopropyl)-'-ethylcarbodimide hydrochloride (EDC) activation. The fluorescent probe was characterized by dynamic light scattering (DLS). The better bioactivity and stability of the fluorescent probe was obtained under the pH value of 8.0, the molecule molar ratio of EDC (1 : 2000) and anti-FLE monoclonal antibodies (1 : 10). The control line (C line) and test line (T line) of a nitrocellulose (NC) filter membrane were sprayed with SPA (0.05 mg mL) and FLE-OVA (1.4 mg mL) solutions with optimal concentration, respectively. A novel method of fluorescent immunochromatographic assay based on quantum dots (QDs-ICA) in this work exhibited good accuracy, reproductivity and excellent specificity under the optimal experimental conditions. Compared with the traditional method for the visual detection of FLE, the developed QDs-ICA can successfully determine FLE residues in pork meat with a better cut-off value of 2.5 ng mL. The QDs-ICA could be adapted for the rapid preliminary detection of FLE residues in pork meat for the first time.
氟罗沙星(FLE)是一种广谱喹诺酮类抗生素,广泛应用于畜牧业、兽医学和水产养殖。食用含有FLE残留的动物源性食品会导致过敏、中毒或耐药性。采用水溶性量子点(CdSe/ZnS)和抗FLE单克隆抗体(mAb),通过N-(3-二甲氨基丙基)-N'-乙基碳二亚胺盐酸盐(EDC)活化法制备荧光探针。通过动态光散射(DLS)对荧光探针进行表征。在pH值为8.0、EDC分子摩尔比为1∶2000和抗FLE单克隆抗体分子摩尔比为1∶10的条件下,荧光探针具有较好的生物活性和稳定性。在硝酸纤维素(NC)滤膜的控制线(C线)和测试线(T线)上分别喷涂最佳浓度的SPA(0.05 mg/mL)和FLE-OVA(1.4 mg/mL)溶液。本文建立的一种基于量子点的荧光免疫层析分析新方法(QDs-ICA)在最佳实验条件下具有良好的准确性、重复性和优异的特异性。与传统的FLE目视检测方法相比,所开发的QDs-ICA能够成功测定猪肉中的FLE残留,其最佳截断值为2.5 ng/mL。QDs-ICA首次可适用于猪肉中FLE残留的快速初步检测。