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基于具有红色发射的上转换纳米扩增子的准确灵敏的微小RNA双响应荧光检测

Accurate and sensitive dual-response fluorescence detection of microRNAs based on an upconversion nanoamplicon with red emission.

作者信息

Huang Lingling, Zhou Yi, Xu Liang, Ruan Xin, Huang Zhao, Ke Yue, Lin Lisong, Tang Qiuling

机构信息

Department of Stomatology, The First Affiliated Hospital of Fujian Medical University Fuzhou 350001 China

Department of Stomatology, National Regional Medical Center, Binhai Campus of the First Affiliated Hospital, Fujian Medical University Fuzhou 350001 China.

出版信息

RSC Adv. 2024 Oct 18;14(45):32911-32921. doi: 10.1039/d4ra05061d. eCollection 2024 Oct 17.

Abstract

Oral squamous cell carcinoma (OSCC) is the most common type of oral cancer. In recent years, researchers have found a close relationship between microRNAs (miRNAs) and OSCC. In addition, miRNAs are highly stable in tissues and circulation, and are also considered potential biomarkers for cancer detection and prognosis. Among a variety of tools for miRNAs with low abundance, single red-emitting UCNP-based biosensors have attracted special interest due to their unique properties, including deep organizational penetration, weak radiation damage, and low autofluorescence. Additionally, the measurement of low-abundance analytes enzyme-free signal amplification is also an effective means. Herein, by taking advantage of red-emitting UCNPs and an enzyme-toehold-mediated strand displacement cascade, a dual-signal amplification biosensor was constructed. The recycled miRNA can be regarded as a catalyst for the assembly of multiple H1/H2 duplexes, which promoted the response signal of augmented analyte expression. Moreover, the proposed biosensors improved the measurement accuracy a dual-signal response to obviously avert false-positive signals. The proposed method was applied to measure miRNA-222 (a model analyte) in serum samples, and the results were similar to those of polymerase chain reaction (PCR), with spiked recoveries ranging from 91.2% to 101.7%. The proposed assay has the merits of high sensitivity, strong recognition, and low background, indicating broad potential for the measurement of diverse analytes in biological samples.

摘要

口腔鳞状细胞癌(OSCC)是最常见的口腔癌类型。近年来,研究人员发现微小RNA(miRNA)与OSCC之间存在密切关系。此外,miRNA在组织和循环中高度稳定,也被认为是癌症检测和预后的潜在生物标志物。在多种用于检测低丰度miRNA的工具中,基于单发射红色上转换纳米粒子(UCNP)的生物传感器因其独特的性质而备受关注,这些性质包括深层组织穿透性、低辐射损伤和低自发荧光。此外,采用无酶信号放大来检测低丰度分析物也是一种有效的手段。在此,利用发射红色荧光的UCNP和酶介导的链置换级联反应,构建了一种双信号放大生物传感器。循环利用的miRNA可被视为组装多个H1/H2双链体的催化剂,从而增强了分析物表达增加时的响应信号。此外,所提出的生物传感器通过双信号响应提高了测量准确性,明显避免了假阳性信号。将所提出的方法应用于血清样本中miRNA-222(一种模型分析物)的检测,结果与聚合酶链反应(PCR)的结果相似,加标回收率在91.2%至101.7%之间。所提出的检测方法具有高灵敏度、强识别能力和低背景的优点,表明在生物样本中检测多种分析物具有广阔的潜力。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5fad/11487471/acd26db54dc4/d4ra05061d-f1.jpg

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