基于酶剪切策略的疏水纸基SERS传感器用于快速和高灵敏检测胃癌中miR-21  

A hydrophobic paper-based SERS sensor based on enzyme cleavage strategy for rapid and highly sensitive detection of miR-21 in gastric cancer

作　　者：张冬[1] 陈淼 曹小卫 王彪[1] 卜迟文[1] 谢峰 陈宇华 刘永霞 ZHANG Dong;CHEN Miao;CAO Xiaowei;WANG Biao;BU Chiwen;XIE Feng;CHEN Yuhua;LIU Yongxia(Guanyun County People’s Hospital,General Surgery Department,222299,China;Tongzhou Hospital of Traditional Chinese Medicine,Department of Gastroenterology,226300,China;Yangzhou University,Medical College,225001,China)

机构地区：[1]灌云县人民医院普外科,222299 [2]通州区中医院消化内科,226300 [3]扬州大学医学院,225001

出　　处：《光散射学报》2025年第1期8-17,共10页The Journal of Light Scattering

基　　金：连云港市卫生健康面上科技项目(202341);灌云县人民医院科研项目(202309);南通市基础研究项目(JC22022033);国家中医药管理局重点研究所开放项目(202256)。

摘　　要：基于AuNCs疏水纸质基底和酶剪切策略,建立了一种快速、高灵敏、特异性检测胃癌(GC)相关miRNA标志物的方法。首先,将金纳米笼(AuNCs)组装到疏水特性的离型纸表面,形成“热点”丰富的阵列式AuNCs基底。接着在阵列式AuNCs基底表面修饰miR-21互补的单链核苷酸ssDNA-21,然后连接4-MBA标记的金银纳米梭(Au-AgNSs),形成Au-AgNSs@4-MBA@ssDNA-21@AuNCs基底的复合结构。最后在ssDNA-21与miR-21杂交后使用双链特异性剪切酶(DSN)选择性剪切ssDNA-21-miR-21双链中DNA磷酸二酯键,进行SERS检测。该SERS微流控芯片检测miR-21的线性范围为1 fM~1 nM,检测限(LOD)0.34 fM。此外,该SERS传感器操作简单,整个测试过程仅30分钟,并且展现出良好的特异性和重现性。使用该SERS传感器成功检测和区分了健康人和GC患者血清中miR-21表达水平的差异。实时定量聚合酶链反应(qRT-PCR)证实了SERS的准确性。Based on the AuNCs hydrophobic paper substrate and enzymatic cleavage strategy,a rapid,highly sensitive and specific method for detecting gastric cancer(GC)-associated miRNA markers was developed.First,gold nanocages(AuNCs)were assembled on the hydrophobic release paper surface to form a Array style AuNCs substrate with rich"hotspots".Then,miR-21 complementary single-stranded nucleotide ssDNA-21 is modified on the surface of the arrayed AuNCs substrate,followed by ligation of the 4-MBA-labled Au–Ag nano-shuttles(Au-AgNSs)to form a complex structure of the Au-AgNSs@4-MBA@ssDNA-21@AuNCs substrate.Finally,after hybridization between ssDNA-21 and miR-21,DNA phosphodiester bonds in ssDNA-21 miR-21 double stranded DNA were selectively cleaved using double stranded specific cleavage enzyme(DSN)for SERS detection.The linear range of the SERS microfluidic chip for detecting miR-21 is 1 fM~1 nM,with a detection limit(LOD)of 0.34 fM.In addition,the SERS sensor is easy to operate,with a testing process of only 30 minutes,and exhibits good specificity and reproducibility.By using this SERS sensor,we successfully detected and distinguished the difference of miR-21 expression levels in serum of healthy people and GC patients.The accuracy of SERS was validated by real-time quantitative polymerase chain reaction(qRT PCR).

关 键 词：表面增强拉曼散射 疏水基底 酶剪切策略 miRNA 胃癌 

分 类 号：O433[机械工程—光学工程] R735.2[理学—光学]