基于Fe_(3)O_(4)@AuNPs富集分离和酶剪切策略的表面增强拉曼光谱传感器用于帕金森病诊断  

Surface-enhanced Raman scattering sensor based on Fe_(3)O_(4)@AuNPs enrichment,separation,and enzymatic shearing strategy for the diagnosis of Parkinson's disease

作　　者：周明[1] 庄艳雯 曹小卫 朱环宇 杜守云 刘敏华[2] Ming Zhou;Yanwen Zhuang;Xiaowei Cao;Huanyu Zhu;Shouyun Du;Minhua Liu(Department of Neurology,Guanyun People's Hospital,Guanyun 222000,China;Department of Critical Care Medicine,Guanyun People's Hospital,Guanyun 222000,China;College of Medicine,Yangzhou University,Yangzhou 225009,China)

机构地区：[1]灌云县人民医院神经内科,灌云222200 [2]灌云县人民医院重症医学科,灌云222200 [3]扬州大学医学院,扬州225009

出　　处：《中国科学:化学》2025年第2期521-529,共9页SCIENTIA SINICA Chimica

基　　金：国家自然科学基金(编号:81701825);连云港市卫生健康面上科技项目(编号:202231);连云港市老龄健康科研项目(编号:L202201)资助项目。

摘　　要：生物标志物的灵敏准确的检测有助于帕金森病(PD)的诊断.本研究制备了一种基于Fe_(3)O_(4)@AuNPs富集分离和酶剪切策略的表面增强拉曼散射(SERS)传感器,用于检测PD血清中miR-221.合成的Fe_(3)O_(4)@AuNPs兼具磁性纳米粒子和金纳米粒子的优点,可以形成大量的“热点”,显著提升了SERS传感器的性能.当miR-221存在时,miR-221与拉曼信号分子Cy5标记的互补单链DNA(ssDNA-221)配对杂交形成miR-221-ssDNA-221双链.在双链特异性剪切酶(DSN)的作用下,核酸双链被切割,导致Cy5远离Fe_(3)O_(4)@AuNPs表面,SERS信号降低.根据Cy5的SERS强度,实现目标miR-221的快速定量检测.结果显示,SERS传感器对血清中miR-221的检测限(LOD)低至5.72aM.此外,该测试平台也具有良好的特异性和重现性.为了验证其临床应用的可行性,使用SERS传感器对PD小鼠和健康小鼠血清中miR-221进行分析,并通过qRT-PCR验证了SERS的准确性.SERS有望成为临床中PD筛查的可靠工具.Sensitive and accurate detection of biomarkers facilitates the diagnosis of Parkinson's disease(PD).In this study,a surface-enhanced Raman scattering(SERS)sensor based on Fe_(3)O_(4)@AuNPs enrichment,separation,and enzymatic shearing strategy was prepared for the detection of miR-221 in PD serum.The synthesized Fe_(3)O_(4)@AuNPs combine the advantages of magnetic nanoparticles and gold nanoparticles,which can form a large number of“hotspots”and thus significantly enhance the performance of the SERS sensor.When miR-221 was present,miR-221 hybridized with complementary single-stranded DNA(ssDNA-221)labeled with the Raman signaling molecule Cy5 to form the miR-221-ssDNA-221 double strand.In the presence of duplex-specific nuclease(DSN),the double-stranded nucleic acid was cleaved,resulting in Cy5 moving away from the surface of Fe_(3)O_(4)@AuNPs and a decrease in the SERS signal.According to the SERS intensity of Cy5,rapid and quantitative detection of target miR-221 was realized.The results showed that the limit of detection(LOD)of the SERS sensor for miR-221 in serum was 5.72 aM.In addition,the test platform also had good specificity and reproducibility.To verify the feasibility of its clinical application,miR-221 was analyzed in serum from PD mice and healthy mice using the SERS sensor,and the accuracy of SERS was verified by qRT-PCR.SERS is expected to be a reliable tool for PD screening in the clinic.

关 键 词：表面增强拉曼散射 磁性富集分离 酶剪切策略 帕金森病 MIRNA 

分 类 号：R742.5[医药卫生—神经病学与精神病学] O657.37[医药卫生—临床医学]