纳米酶增强免疫层析法定量检测血液中的降钙素原  

Quantitative Detection of Procalcitonin in Blood by Nanozyme-based Lateral Flow Immunoassay

作　　者：郑玥 林童 熊勇华[1,2] 许梦硕 严喜鸾 郭徐静 杨磊 郭亮[1,2] ZHENG Yue;LIN Tong;XIONG Yong-Hua;XU Meng-Shuo;YAN Xi-Luan;GUO Xu-Jing;YANG Lei;GUO Liang(State Key Laboratory of Food Science and Resources,Nanchang University,Nanchang 330047,China;Jiangxi-OAI Joint Research Institute,Nanchang University,Nanchang 330047,China;School of Pharmacy,Nanchang University,Nanchang 330006,China)

机构地区：[1]南昌大学,食品科学与资源挖掘全国重点实验室,南昌330047 [2]南昌大学中德联合研究院,南昌330047 [3]南昌大学药学院,南昌330006

出　　处：《分析化学》2024年第8期1082-1093,共12页Chinese Journal of Analytical Chemistry

基　　金：江西省自然科学基金项目(No.20232BAB205087);国家自然科学基金项目(No.82360857)资助。

摘　　要：建立了一种以金、铂纳米粒子修饰的金属有机框架材料(MAP)为标记物的降钙素原(Procalcitonin,PCT)快速定量免疫层析检测方法。本方法以静电吸附法制备的MAP-抗PCT单克隆抗体偶联物为检测探针,将抗PCT多克隆抗体和羊抗鼠IgG分别喷涂硝酸纤维素(NC)膜构建试纸条的检测线和质控线,采用基于信号放大的夹心免疫法定量检测PCT。结果表明,MAP免疫层析试纸条具有灵敏度高、特异性强和稳定性好等优点,利用其检测PCT的动态检测范围为0.61 pg/mL~320 ng/mL,检出限为0.25 pg/mL,日内和日间精密度(RSD)均小于15%。实际样品检测结果表明,本方法仅需微量样本即可检测全血中PCT的浓度,对于炎症反应的早期诊断、监测治疗及预后判断具有重要意义。A rapid quantitative immunochromatographic assay for procalcitonin(PCT)using metal-organic frameworks modified with gold and platinum nanoparticles(MAPs)as labels was established in this work.The detection probe was prepared by conjugating MAPs with anti-PCT monoclonal antibody via an electrostatic adsorption method.Anti-PCT polyclonal antibody and sheep anti-mouse IgG were sprayed onto the nitrocellulose(NC)membrane as the test line and quality control line,respectively,to construct immunochromatographic strip for PCT quantitative detection via signal-amplification-based sandwich immunoassay.The results showed that the MAP-based immunochromatographic test had high sensitivity,high specificity,and good stability.The dynamic range for detection of PCT was 0.61 pg/mL–320 ng/mL,the detection limit was 0.25 pg/mL,and the intra-day and inter-day precision(Relative standard deviation)were less than 15%.The results of real sample analysis showed that a quite low volume of sample was required for detection of PCT in whole blood,which was of great significance for the early diagnosis,monitoring and treatment,and prognosis of inflammation.

关 键 词：侧流免疫层析法 降钙素原 快速检测 纳米酶 

分 类 号：O657[理学—分析化学] R446.6[理学—化学]