Aptamer-based Sandwich Assay and its Clinical Outlooks for Detecting Lipocalin-2 in Hepatocellular Carcinoma (HCC).
Aptamer-based Sandwich Assay and its Clinical Outlooks for Detecting Lipocalin-2 in Hepatocellular Carcinoma (HCC).
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DOI：
10.1038/srep10897
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发表时间：
2015-06-03
影响因子：
4.6
通讯作者：
Kim YH
中科院分区：
文献类型：
--
作者：
Lee KA;Ahn JY;Lee SH;Singh Sekhon S;Kim DG;Min J;Kim YH
我们验证了一种基于DNA适配子的单链诊断方法，该方法能够在夹心法中检测临床相关的肝细胞癌患者血清中的Lipocalin-2(Lcn2)。用SELEX方法筛选出Lcn2的9个适配子(Lcn2_apta1~Lcn2_apta9)，并通过表面等离子体共振(SPR)和斑点印迹分析筛选出一对三明治对(Lcn2_apta2和Lcn2_apta4)。该适配子夹心构建的结果表明，LCN2在2.5~5 0 0 ng m L−1的浓度范围内被灵敏地检测到，检测下限为0.6 ng m L−1。适体夹心法显示了在临床相关血清水平下Lcn2的良好动态范围，覆盖了每毫升亚纳克浓度。这一新方法为检测低丰度和中等丰度的血清生物标志物提供了一种简单而稳健的方法。它由功能化、杂交和信号读出组成，不需要稀释。研究结果证明了适配子夹心分析平台诊断肝细胞癌的能力，以及它对护理点检测(POCT)系统的潜在适用性。
We validated a single-stranded, DNA aptamer-based, diagnostic method capable of detecting Lipocalin-2 (LCN2), a biomarker from clinically relevant hepatocellular carcinoma (HCC) patient serum, in the sandwich assay format. Nine aptamers (LCN2_apta1 to LCN2_apta9) for LCN2 were screened with SELEX processes, and a sandwich pair (LCN2_apta2 and LCN2_apta4) was finally chosen using surface plasmon resonance (SPR) and dot blotting analysis. The result of the proposed aptamer sandwich construction shows that LCN2 was sensitively detected in the concentration range of 2.5–500 ng mL−1 with a limit of detection of 0.6 ng mL−1. Quantitative measurement tests in HCC patients were run on straight serum and were compared with the performance of the conventional antibody-based ELISA kit. The aptamer sandwich assay demonstrated an excellent dynamic range for LCN2 at clinically relevant serum levels, covering sub-nanogram per mL concentrations. The new approach offers a simple and robust method for detecting serum biomarkers that have low and moderate abundance. It consists of functionalization, hybridization and signal read-out, and no dilution is required. The results of the study demonstrate the capability of the aptamer sandwich assay platform for diagnosing HCC and its potential applicability to the point-of-care testing (POCT) system.
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影响因子：
1.8
作者：
Chiappini F
通讯作者：
Chiappini F
影响因子：
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作者：
Ambrosi, Adriano;Airo, Federico;Merkoci, Arben
通讯作者：
Merkoci, Arben
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作者：
Howard, BA;Wang, MZ;Patz, EF
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Patz, EF
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作者：
Lee, Seung-Pyo;Park, Kyung Woo;Kim, Hyo-Soo
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Kim, Hyo-Soo
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作者：
Gupta D;Lis CG
通讯作者：
Lis CG