School of Food and Biological Engineering, Jiangsu University, Zhenjiang, Jiangsu, China.
Jiangsu Meizheng Biological Technology Co. Ltd., Wuxi, Jiangsu, China.
J Environ Sci Health B. 2022;57(7):517-525. doi: 10.1080/03601234.2022.2068908. Epub 2022 Apr 27.
A rapid colloidal gold immunochromatography assay (GICA) for the detection of pefloxacin (PEF) was established and optimized. The anti-PEF monoclonal antibody (mAb) was used to target PEF as a colloidal gold-mAb conjugate. The mAb belonged to the IgG2b subtype, lambda light chain, the affinity constant (Ka) was 5.21 × 10 L·mol, and its half maximal inhibitory concentration (IC) was 0.23 ng·mL. No obvious cross-reactivity (CR) was observed with other common fluoroquinolone antibiotics, including ciprofloxacin (CIP), norfloxacin (NOR), lomefloxacin (LOM) and ofloxacin (OFL). The visual limit of detection (vLOD) of the optimized GICA was 2 ng·g under the conventional pretreatment method, and the assay was completed in 15 min. Liquid chromatography tandem-mass spectrometry (LC-MS/MS) was employed to confirm the performance of the strip. In addition, a novel pretreatment was established and compared with conventional pretreatment. Without the removal of organic solvents, the novel pretreatment method reduced the sample pretreatment time (more than 10 min). The vLOD of the optimized GICA was also 2 ng·g when applying the novel pretreatment method. In conclusion, the proposed PEF-GICA could detect samples containing PEF rapidly and accurately, and the novel pretreatment method saved the time of sample pretreatment and improved the efficiency of detection.
建立并优化了一种用于检测氟甲喹（PEF）的胶体金免疫层析检测法（GICA）。以抗氟甲喹单克隆抗体（mAb）作为胶体金-mAb 结合物来靶向氟甲喹。该 mAb 属于 IgG2b 亚型，λ 轻链，亲和常数（Ka）为 5.21×10 L·mol，半抑制浓度（IC）为 0.23 ng·mL。与其他常见的氟喹诺酮类抗生素（如环丙沙星（CIP）、诺氟沙星（NOR）、洛美沙星（LOM）和氧氟沙星（OFL））没有明显的交叉反应（CR）。在常规预处理方法下，优化后的 GICA 的目视检测限（vLOD）为 2ng·g，检测时间为 15min。采用液相色谱串联质谱法（LC-MS/MS）对条带进行了性能确认。此外，还建立了一种新的预处理方法，并与常规预处理方法进行了比较。在不去除有机溶剂的情况下，新的预处理方法减少了样品预处理时间（超过 10min）。采用新的预处理方法，优化后的 GICA 的 vLOD 也为 2ng·g。总之，所提出的 PEF-GICA 可快速准确地检测含氟甲喹的样品，新的预处理方法节省了样品预处理时间，提高了检测效率。
