基于量子点的荧光免疫层析法分析猪饲料、肌肉和肝脏样品中的维吉尼霉素M1

Jingming Zhou, Wenjing Qian, Qingbao Yang, Chao Liang, Yumei Chen, Aiping Wang, Gaiping Zhang
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引用次数: 2

摘要

基于virginiamycin M1 (VIR M1)高敏感特异性单克隆抗体(mAb),首次建立了基于量子点的VIR M1荧光免疫层析(QDs-ICA)快速、灵敏分析方法。当用于酶联免疫吸附试验(ELISA)时,该单抗对其他三种类似物的半最大抑制浓度(IC50)为0.5 ng/mL,交叉反应性(CR)值低于0.1%。该单抗与ZnCdSe/ZnS(核/壳)量子点偶联,最大发射波长为610 nm(橙红色),作为荧光探针,以提高QDs- ica灵敏度。QDs-ICA的截止值为12.5 ng/mL。QDs-ICA在0.7 ~ 14.5 ng/mL范围内呈线性关系,定量限为0.7 ng/mL。与现有的VIR M1分析方法相比,QDs-ICA具有更高的灵敏度。对猪饲料、肌肉和肝脏样品中添加的VIR M1浓度进行分析,检测内回收率为94.0% ~ 111.6%,变异系数(CV)最高为6.7%;检测间回收率为94.7% ~ 107.6%,变异系数最高为9.4%。综上所述,QDs-ICA可作为分析动物饲料和动物源性食品中VIR M1的一种潜在方法。
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Analysis of virginiamycin M1 in swine feed, muscle and liver samples by quantum dots-based fluorescent immunochromatographic assay
Abstract Based on a highly sensitive and specific monoclonal antibody (mAb) against virginiamycin M1 (VIR M1), a quantum dots-based fluorescent immunochromatographic assay (QDs-ICA) for quick and sensitive analysis of VIR M1 was established for the first time. The mAb showed a half-maximal inhibitory concentration (IC50) of 0.5 ng/mL and cross-reactivity (CR) values below 0.1% for other three analogues when used in enzyme-linked immunosorbent assay (ELISA). The mAb was conjugated to ZnCdSe/ZnS (core/shell) QDs with maximum emission wavelength of 610 nm (orange-red) which was selected as fluorescent probe to increase QDs-ICA sensitivity. The cut-off value of QDs-ICA was 12.5 ng/mL. QDs-ICA showed a linear range from 0.7 to 14.5 ng/mL with a limit of quantification of 0.7 ng/mL. Compared with existing methods for the analysis of VIR M1, the QDs-ICA exhibited higher sensitivity. For analysis of VIR M1 concentrations spiked into swine feed, muscle and liver samples, recovery rates ranged from 94.0% to 111.6% with the highest coefficient of variation (CV) of 6.7% for intra-assay, and for inter-assay ranged from 94.7% to 107.6% with the highest CV of 9.4%. In conclusion, the QDs-ICA could be a potential method for analyzing VIR M1 in animal feed and animal-derived food.
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