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基于铈元素(Ce)的荧光侧向液流免疫层析(lateral-flow immunoassays,LFIA)技术,开发一种用时短、检测限低的方法,定量测定降钙素原(procalcitonin,PCT),以满足临床检测和诊断的需求。经反向微乳化、氨基修饰、包被Ce,合成Ce微球;其经活化、洗涤,用PCT抗体标记结合和封闭制备微球悬浮剂,固化液相、固定包被和质控抗体、组装和切割等步骤制备PCT试纸条,检测PCT标准品;通过精密度、正确度、回收率、交叉反应和热稳定实验分析该PCT试纸条的临床价值。实验结果显示:用1%含量的Ce微球、100μg标记抗体、Blockmaster TM CE510封闭剂制备的PCT试纸条,采用LFIA法检测PCT,仅需4 min完成检测,大大缩短实验时间;检测限达0.10μg/L;且无边缘和钩状效应、交叉反应;成本低、精密度和准确度高、回收率佳、热稳定性好。因此,本实验基于Ce的LFIA法,成功研发了一种快速、灵敏的PCT床旁定量检测。
Based on the fluorescence lateral liquid flow immunoassay (CeFIA) technique, we developed a method for the quantitative determination of procalcitonin (PCT) with short time and low detection limit Meet the needs of clinical testing and diagnosis. After reverse microemulsification, amino modification, coating Ce, synthesis of Ce microspheres; its activation, washing, PCT antibody labeling and microspheres suspension preparation suspension, the liquid phase was fixed, fixed coating and quality control antibody assembly And cutting steps to prepare PCT test strips, PCT standards were tested; the clinical value of PCT test strips was analyzed by precision, accuracy, recovery, cross-reaction and thermal stability experiments. The experimental results showed that using PCT strip prepared with 1% Ce microspheres, 100μg labeled antibody and Blockmaster TM CE510 blocker, PCT was detected by LFIA method and the test was only completed in 4 min, which greatly shortened the experimental time. The limit of detection 0.10μg / L; and no edge and hook effect, cross-reaction; low cost, precision and accuracy, good recovery, good thermal stability. Therefore, based on the Ce-based LFIA method, this experiment successfully developed a fast and sensitive quantitative detection of PCT bedside.