Background

Alpha-fetoprotein (AFP) has been used as a biomarker for early diagnosis of hepatocellular carcinoma (HCC) for a long time. Aptamer is a type of material that is receiving attention as a new alternative in the diagnosis and treatment of diseases.

Objective

We applied aptamer to AFP detection and utilized in silico analysis techniques. Here, we describe the selection of aptamers that can bind to AFP as well as in vitro and in silico validation of the binding site. We also developed an aptamer-based detection platform called hybrid-aptablotting sandwich assay (Hybrid-ABSA) and a point-of-care AFP detection kit.

Results

To construct the AFP detection platform, we screened a total of 18 candidate AFP-binding aptamers and identified a pair of aptamers without non-overlapping-binding sites using dot blotting. We validated results of dot blotting using in silico 3D binding structure docking simulation analysis. The aptamer-based Hybris-ABSA for AFP detection using this pair of aptamers demonstrated superior detection performance to antibody-based ELISA detection methods. Furthermore, we confirmed that AFP could be detected using the developed point-of-care lateral flow In Vitro Diagnostic (IVD) kit.

Conclusion

Results of this study demonstrate that aptamer-based disease diagnostic technology has potential of replace conventional diagnostic methods. This study also confirms that in silico techniques have sufficient utility in biological assays.

中文翻译：

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适体3D结构分析及肝细胞癌诊断平台

背景

甲胎蛋白（AFP）长期以来一直被用作肝细胞癌（HCC）早期诊断的生物标志物。适体是一种作为疾病诊断和治疗的新替代品而受到关注的材料。

客观的

我们将适体应用于 AFP 检测并用于计算机分析技术。在这里，我们描述了可以与 AFP 结合的适体的选择以及结合位点的体外和计算机验证。我们还开发了一种基于适体的检测平台，称为混合适体印迹夹心测定（Hybrid-ABSA）和护理点 AFP 检测试剂盒。

结果

为了构建AFP检测平台，我们总共筛选了18个候选AFP结合适体，并使用斑点印迹法鉴定了一对没有非重叠结合位点的适体。我们使用计算机 3D 结合结构对接模拟分析验证了点印迹的结果。使用这对适体进行 AFP 检测的基于适体的 Hybris-ABSA 表现出优于基于抗体的 ELISA 检测方法的检测性能。此外，我们证实可以使用开发的护理点侧流体外诊断 (IVD) 试剂盒检测 AFP。

结论

本研究结果表明，基于适配体的疾病诊断技术具有取代传统诊断方法的潜力。这项研究还证实了计算机技术在生物测定中具有足够的实用性。