Sensitive immunoassays based on a monoclonal antibody for detection of marbofloxacin in milk
| Autor: | Keyu Xing, Zhen Guo, Juan Peng, Meixia Luo, Debin Guo, Weihua Lai |
|---|---|
| Rok vydání: | 2020 |
| Předmět: |
Fleroxacin
Danofloxacin Enzyme-Linked Immunosorbent Assay Gold Colloid Cross Reactions Pefloxacin Mice Marbofloxacin Genetics Enrofloxacin medicine Animals Veterinary drug Immunoassay Mice Inbred BALB C Chromatography Chemistry Antibodies Monoclonal Drug Residues Anti-Bacterial Agents Milk Lomefloxacin Cattle Female Animal Science and Zoology Ofloxacin Fluoroquinolones circulatory and respiratory physiology Food Science medicine.drug |
| Zdroj: | Journal of Dairy Science. 103:7791-7800 |
| ISSN: | 0022-0302 |
| Popis: | Marbofloxacin (MBF) is a key class of synthetic fluoroquinolone antibiotic that is commonly used as a veterinary drug. However, excess residue of MBF in animal-derived food samples, such as milk, is harmful to consumers. In this study, 4 mAb against MBF, namely, M4E3, M7A6, M3C7, and M5C6, were produced. Indirect competitive (ic) ELISA revealed that the M4E3 exhibited the highest sensitivity with a half-maximal inhibitory concentration (IC50) of 0.07 ng/mL and a limit of detection of 0.01 ng/mL for detection of MBF. The results of the cross-reactivity experiment revealed that the M4E3 could detect lomefloxacin, ofloxacin, fleroxacin, pefloxacin, danofloxacin, and enrofloxacin sensitively with IC50 of 0.02, 0.07, 0.18, 0.27, 0.30, and 0.32 ng/mL, respectively. Meanwhile, a cross-reactivity experiment showed that the M4E3 had a crossover rate of more than 20% with these fluoroquinolone analogs. A weak crossover rate below 1.11% was observed with 14 other fluoroquinolone analogs. The recovery rate of MBF in milk ranged from 72.28% to 129.19%, which showed that the developed indirect competitive ELISA was effective in detecting MBF in milk. Furthermore, a visual colloidal gold-based immunochromatographic assay was developed for detecting MBF with a cut-off value of 1 ng/mL in both phosphate-buffered saline and a milk sample by using this mAb. Given this sensitive mAb, both indirect competitive ELISA and colloidal gold-based immunochromatographic assay could be effective screening tools for detection of MBF in milk. |
| Databáze: | OpenAIRE |
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