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In Situ Exosomal MicroRNA Determination by Target-Triggered SERS and Fe3O4@TiO2-Based Exosome Accumulation
ACS Sensors ( IF 8.2 ) Pub Date : 2021-02-08 , DOI: 10.1021/acssensors.0c01900 Shuqin Jiang 1 , Qing Li 1 , Chongwen Wang 2 , Yuanfeng Pang 1 , Zhiwei Sun 1 , Rui Xiao 2
ACS Sensors ( IF 8.2 ) Pub Date : 2021-02-08 , DOI: 10.1021/acssensors.0c01900 Shuqin Jiang 1 , Qing Li 1 , Chongwen Wang 2 , Yuanfeng Pang 1 , Zhiwei Sun 1 , Rui Xiao 2
Affiliation
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Exosomal microRNAs (miRNAs) have been proved to be important biomarkers for the early diagnosis of cancers. However, the accurate quantification of exosomal miRNAs is hampered either by laborious exosome isolation and lysis or by RNA extraction and the amplification process. Here, we reported an in situ platform for direct exosomal miRNAs from serum samples. First, locked nucleic acid (LNA)-modified Au@DTNB (DTNB is the Raman reporter molecule 5,5′-dithiobis-(2-nitrobenzoic acid)) was synthesized as surface-enhanced Raman scattering (SERS) tags to enter into exosomes and assemble with target miRNAs to induce hot-spot SERS signals. Second, Fe3O4@TiO2 nanoparticles were added to enrich the exosomes through affinity interaction of the TiO2 shell for further SERS detection. Based on the platform, target miRNAs can be directly qualified in situ with a detection limit of 0.21 fM, which is better or comparable with quantitative reverse transcription polymerase chain reaction (qRT-PCR) and other in situ methods reported before. Moreover, neither capture antibody nor ultracentrifugation pretreatment was needed in the whole detection procedure. Using exosomal miRNA-10b as a proof of concept, pancreatic ductal adenocarcinoma (PDAC) patients can be recognized from normal controls (NCs) with an accuracy of 99.6%. The simple and sensitive in situ exosomal miRNA detection assay can be seen as a noninvasive liquid biopsy assay for clinical cancer diagnostic adaption.
中文翻译：
目标触发的SERS和基于Fe 3 O 4 @TiO 2的外来体累积的原位外泌微RNA测定
外泌体微小RNA（miRNA）已被证明是癌症早期诊断的重要生物标志物。但是，费力的外泌体分离和裂解或RNA提取和扩增过程阻碍了外泌体miRNA的准确定量。在这里，我们报道了来自血清样品的直接外泌体miRNA的原位平台。首先，合成锁核酸（LNA）修饰的Au @ DTNB（DTNB是拉曼报告分子5,5'-二硫代双-（2-硝基苯甲酸））作为表面增强拉曼散射（SERS）标签进入外泌体并与目标miRNA组装以诱导热点SERS信号。其次，添加Fe 3 O 4 @TiO 2纳米颗粒以通过TiO 2的亲和相互作用富集外泌体外壳用于进一步的SERS检测。基于该平台，目标miRNA可以直接原位鉴定，检出限为0.21 fM，与定量逆转录聚合酶链反应（qRT-PCR）和之前报道的其他原位方法相比更好或更可比。此外，在整个检测过程中都不需要捕获抗体或超速离心预处理。使用外泌体miRNA-10b作为概念验证，可以从正常对照（NCs）中识别出胰腺导管腺癌（PDAC）患者，其准确率为99.6％。简单而灵敏的原位外泌体miRNA检测方法可以看作是一种用于临床癌症诊断适应性的无创液体活检方法。
更新日期：2021-03-26
中文翻译：
目标触发的SERS和基于Fe 3 O 4 @TiO 2的外来体累积的原位外泌微RNA测定
外泌体微小RNA（miRNA）已被证明是癌症早期诊断的重要生物标志物。但是，费力的外泌体分离和裂解或RNA提取和扩增过程阻碍了外泌体miRNA的准确定量。在这里，我们报道了来自血清样品的直接外泌体miRNA的原位平台。首先，合成锁核酸（LNA）修饰的Au @ DTNB（DTNB是拉曼报告分子5,5'-二硫代双-（2-硝基苯甲酸））作为表面增强拉曼散射（SERS）标签进入外泌体并与目标miRNA组装以诱导热点SERS信号。其次，添加Fe 3 O 4 @TiO 2纳米颗粒以通过TiO 2的亲和相互作用富集外泌体外壳用于进一步的SERS检测。基于该平台，目标miRNA可以直接原位鉴定，检出限为0.21 fM，与定量逆转录聚合酶链反应（qRT-PCR）和之前报道的其他原位方法相比更好或更可比。此外，在整个检测过程中都不需要捕获抗体或超速离心预处理。使用外泌体miRNA-10b作为概念验证，可以从正常对照（NCs）中识别出胰腺导管腺癌（PDAC）患者，其准确率为99.6％。简单而灵敏的原位外泌体miRNA检测方法可以看作是一种用于临床癌症诊断适应性的无创液体活检方法。
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