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一种新型的 SERS-侧向流分析(LFA)检测盘,用于监测乳腺癌细胞焦亡过程中的 miR-155-5p。

A novel SERS-lateral flow assay (LFA) tray for monitoring of miR-155-5p during pyroptosis in breast cancer cells.

机构信息

Department of Oncology, The Second Affiliated Hospital of Soochow University, Suzhou, 214122, China.

Department of Oncology, The Affiliated Hospital of Yangzhou University, Yangzhou University, Yangzhou, 225000, China.

出版信息

Anal Methods. 2024 Jun 20;16(24):3878-3894. doi: 10.1039/d4ay00363b.

Abstract

In the study, a novel surface-enhanced Raman scattering (SERS)-lateral flow assay (LFA) tray for the real-time detection of pyroptosis-associated miR-155-5p in breast cancer cells was established and validated. The SERS probe modified with monoclonal antibodies and functionalized HP1@5-FAM was first synthesized. When miR-155-5p was present, HP1@5-FAM on the SERS probe specifically recognized target miRNAs and hybridized with them, resulting in HP2 on the T line only capturing some SERS probes that were not bound to miR-155-5p. The T line appeared as a light orange band or there was no color change, and the corresponding Raman detection result showed a weak or insignificant Raman signal. The SERS probe showed high selectivity, satisfactory stability, and excellent reproducibility, and the limit of detection (LOD) for miR-155-5p was 7.26 aM. Finally, the proposed SERS-LFA tray was applied to detect miR-155-5p in MBA-MD-468 cells that underwent varying degrees of pyroptosis, and the detection results of SERS were consistent with those of the conventional real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR) assay. The study demonstrated that the SERS-LFA tray was a convenient and ultrasensitive method for miR-155-5p real-time detection, which could provide more detailed information for pyroptosis and be of potential value in guiding the treatment of breast cancer.

摘要

在这项研究中,建立并验证了一种用于实时检测乳腺癌细胞中细胞焦亡相关 miR-155-5p 的新型表面增强拉曼散射(SERS)-侧向流分析(LFA)检测盘。首先合成了修饰有单克隆抗体并功能化 HP1@5-FAM 的 SERS 探针。当存在 miR-155-5p 时,SERS 探针上的 HP1@5-FAM 特异性识别靶 miRNAs 并与其杂交,导致 T 线上仅捕获一些未与 miR-155-5p 结合的 SERS 探针。T 线呈现浅橙色带或无颜色变化,相应的拉曼检测结果显示弱或不明显的拉曼信号。SERS 探针表现出高选择性、令人满意的稳定性和出色的重现性,miR-155-5p 的检测限(LOD)为 7.26 aM。最后,将所提出的 SERS-LFA 检测盘应用于检测经历不同程度细胞焦亡的 MBA-MD-468 细胞中的 miR-155-5p,SERS 的检测结果与传统的实时定量逆转录聚合酶链反应(qRT-PCR)检测结果一致。该研究表明,SERS-LFA 检测盘是一种用于实时检测 miR-155-5p 的便捷和超灵敏方法,可为细胞焦亡提供更详细的信息,并可能对指导乳腺癌治疗具有潜在价值。

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