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.
作者
周明
庄艳雯
曹小卫
朱环宇
杜守云
刘敏华
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)
出处
《中国科学:化学》
北大核心
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。
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
surface-enhanced Raman scattering
magnetic enrichment separation
enzymatic shearing strategy
Parkinson's disease
miRNA
