目的：建立一种简便、快速、特异的利奈唑胺血药浓度荧光免疫层析检测法。方法：将脱乙酰利奈唑胺定向合成具有单一羧基结构的利奈唑胺衍生物。采用碳二亚胺法将利奈唑胺衍生物与牛血清蛋白和卵清蛋白分别偶联制备完全抗原。将利奈唑胺-牛血清蛋白偶联物免疫Balb/c小鼠制备单克隆抗体。荧光标记利奈唑胺单抗，将利奈唑胺-卵清蛋白完全抗原和羊抗兔IgG抗体喷涂于硝酸纤维素膜分别作为检测线和质控线，以竞争性免疫抑制反应模式建立了利奈唑胺血药浓度荧光免疫层析检测法。基于以上检测方法建立利奈唑胺的标准曲线，并对该检测法进行方法学评价。并采用HPLC方法测定，对两方法的结果进行比较。结果：通过优化体系，所建立的检测法IC50=5.28μg·mL-1，线性范围为0.78~50 μg·mL-1，灵敏度为0.61 μg·mL-1，精密度均小于10%，回收率为78.4%~123%。与14种与利奈唑胺结构相似以及联合用药相关药物无明显交叉反应。分别检测67份临床利奈唑胺血样与HPLC法对比，发现2种方法相关性较好，R2=0.929，大于0.90。结论：本实验成功建立了一种操作简便、快速、高灵敏度、特异性强的利奈唑胺荧光免疫层析检测方法，为进一步临床血药浓度监测的应用打下了基础。
Objective: To establish a simple, rapid, specific fluoroimmunoassay method for the determination of plasma concentration of linezolid. Methods: Deacetylation linezolid was used to synthesize linezolid derivatives with single carboxyl structure. The complete antigen was prepared by coupling the linezolid derivatives with bovine serum albumin and ovalbumin by carbodiimide method. The monoclonal antibody was obtained by immunizing Balb/c mice with linezolid-bovine serum albumin conjugate, which was labeled with fluorescence subsequently. The linezolid-ovalbumin and goat anti-rabbit antibody IgG were immobilized on the nitrocellulose membrane as the test line and control line, the competitive immunosuppression model was adopted to establish linezolid plasma concentration of fluorescence immunochromatographic assay. Based on the above detection methods, the standard curve of linezolid was established, and the method was validated by methodology. HPLC method was used to compare the results of the two methods. Results: By optimizing the system, we established the assay, of which IC50 was 5.28 μg·mL-1, the linear range was 0.78-50 μg ·mL-1, the sensitivity was 0.61 μg·mL-1, the precision was less than 10%, the recovery rate was 78.4%-123%. No cross-reaction was found in 14 kinds of drugs whose structures were similar to linezolid or usually combined use with linezolid. At last, this assay and HPLC were used to detect 67 clinical linezolid blood samples, the results showed that the correlation between the two methods was better, and the r2=0.929, greater than 0.90. Conclusions: In this study, we successfully established a simple, rapid, high sensitivity and specific immunochromatographic method for the detection of linezolid, which contributed to the foundation for the further application of clinical plasma concentration monitoring.
