Department of Pharmacy, The Second Affiliated Hospital of Zunyi Medical University, Zunyi, 563000, Guizhou Province, China.
College of Pharmaceutical Sciences, Southwest University, Chongqing, 400715, China.
Mikrochim Acta. 2023 Aug 4;190(9):342. doi: 10.1007/s00604-023-05917-6.
As a front-line chemotherapeutic drug for maintenance and consolidation therapy, methotrexate (MTX) has widely been applied to treat various tumors and some inflammatory diseases. However, because of its severe toxicity ascribed to low selectivity, it is necessary to monitor therapeutic drugs in high-dose MTX therapeutic regimens to ensure treatment safety. In this work, we developed a fluorescent immunochromatographic test strip (FITS) for monitoring MTX by employing time-resolved fluorescent microspheres as signal probes. With a competitive immunoassay mode, the FITS for MTX shows a super-wide dynamic range of 10 pM-10 μM, covering the entire clinical therapeutic concentration range of MTX. Therapeutic drug monitoring of MTX can be achieved within 7 min with high specificity, facilitating the timely rescue of drug poisoning led by high-dose MTX treatment. The method was employed for monitoring MTX in the spiked human serum, urine, and milk, showing acceptable recoveries ranging from 94.0 to 110.0%. The established FITS has been applied to MTX detection in serum obtained from high-dose MTX treatment. The results from FITS and enzyme multiplied immunoassay technique showed no significant difference, suggesting its reliability for usage in real biological samples. The device shows promise in point-of-care therapeutic drug monitoring for resource-limited countries and institutes, which significantly facilitates overcoming the lag time between sampling and results.
甲氨蝶呤（MTX）作为一种用于维持和巩固治疗的一线化疗药物，已广泛用于治疗各种肿瘤和一些炎症性疾病。然而，由于其选择性低，毒性严重，因此有必要在高剂量 MTX 治疗方案中监测治疗药物，以确保治疗安全。在这项工作中，我们开发了一种荧光免疫层析测试条（FITS），通过使用时间分辨荧光微球作为信号探针来监测 MTX。采用竞争免疫分析模式，MTX 的 FITS 显示出超宽的动态范围为 10 pM-10 μM，涵盖了 MTX 的整个临床治疗浓度范围。通过该方法可以在 7 分钟内实现 MTX 的治疗药物监测，具有高特异性，有利于及时抢救因高剂量 MTX 治疗引起的药物中毒。该方法已用于人血清、尿液和牛奶中 MTX 的监测，回收率在 94.0%至 110.0%之间，结果令人满意。建立的 FITS 已应用于高剂量 MTX 治疗患者的血清 MTX 检测。FITS 和酶联免疫吸附试验技术的结果没有显著差异，表明其在实际生物样本中使用的可靠性。该设备有望在资源有限的国家和机构中实现即时治疗药物监测，显著缩短了采样和结果之间的时间滞后。
