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Ultrasensitive enzyme-free fluorescent detection of VEGF165 based on target-triggered hybridization chain reaction amplification†
RSC Advances ( IF 3.9 ) Pub Date : 2018-07-19 00:00:00 , DOI: 10.1039/c8ra04721a Qingzhen Zhou 1 , Hongxia Yan 2 , Fengying Ran 1 , Jianjun Cao 1 , Long Chen 1 , Bing Shang 1 , Hao Chen 1 , Jian Wei 1 , Qinhua Chen 1
RSC Advances ( IF 3.9 ) Pub Date : 2018-07-19 00:00:00 , DOI: 10.1039/c8ra04721a Qingzhen Zhou 1 , Hongxia Yan 2 , Fengying Ran 1 , Jianjun Cao 1 , Long Chen 1 , Bing Shang 1 , Hao Chen 1 , Jian Wei 1 , Qinhua Chen 1
Affiliation
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Sensitive detection of vascular endothelial growth factor (VEGF165) is important for early cancer disease diagnosis in the clinic. A sensitive fluorescent sensing platform for VEGF165 detection is developed in this work. It is based on a target-triggered hybridization chain reaction (HCR) and graphene oxide (GO) selective fluorescence quenching. In this assay, in the presence of the VEGF165, the hairpin structure of Hp opens up and the initiation sequence will be exposed to Hp1 to open its hairpin structure. Then the opened Hp1 hybridizes with Hp2 to expose the complementary sequence of Hp1 which hybridizes with Hp1 again by HCR. Thus HCR would be initiated, generating super-long dsDNA. After the HCR, the double strands of the HCR product cannot be adsorbed on the GO surface. As a result, the HCR product gives a strong fluorescence signal which is dependent on the concentration of VEGF165. By using VEGF165 as a model analyte, the assay provides a highly sensitive fluorescence detection method for VEGF165 with a detection limit down to 20 pg mL−1. The proposed aptasensing strategy based on target-triggered HCR amplification can thus be realized. It was successfully applied to the determination of VEGF165 in spiked human serum, urine and saliva. Therefore, it can easily have wide applications in the diagnosis of vital diseases.
中文翻译：
基于靶标触发杂交链式反应扩增的 VEGF165 超灵敏无酶荧光检测†
血管内皮生长因子（VEGF 165）的灵敏检测对于临床早期癌症疾病诊断具有重要意义。这项工作开发了一种用于 VEGF 165检测的灵敏荧光传感平台。它基于靶标触发的杂交链式反应 (HCR) 和氧化石墨烯 (GO) 选择性荧光猝灭。在此测定中，在存在 VEGF 165的情况下，Hp 的发夹结构打开，并且起始序列将暴露于 Hp1 以打开其发夹结构。然后打开的Hp1与Hp2杂交，暴露出Hp1的互补序列，通过HCR再次与Hp1杂交。这样HCR就会被启动，产生超长的双链DNA。HCR后，HCR产物的双链不能吸附在GO表面上。因此，HCR 产物发出强烈的荧光信号，该信号取决于 VEGF 165的浓度。通过使用VEGF 165作为模型分析物，该测定提供了一种高度灵敏的VEGF 165荧光检测方法，检测限低至20 pg mL -1。因此可以实现所提出的基于靶标触发的HCR扩增的适体传感策略。成功应用于人血清、尿液和唾液中VEGF 165的测定。因此，它很容易在重大疾病的诊断中得到广泛的应用。
更新日期：2018-07-19
中文翻译：
基于靶标触发杂交链式反应扩增的 VEGF165 超灵敏无酶荧光检测†
血管内皮生长因子（VEGF 165）的灵敏检测对于临床早期癌症疾病诊断具有重要意义。这项工作开发了一种用于 VEGF 165检测的灵敏荧光传感平台。它基于靶标触发的杂交链式反应 (HCR) 和氧化石墨烯 (GO) 选择性荧光猝灭。在此测定中，在存在 VEGF 165的情况下，Hp 的发夹结构打开，并且起始序列将暴露于 Hp1 以打开其发夹结构。然后打开的Hp1与Hp2杂交，暴露出Hp1的互补序列，通过HCR再次与Hp1杂交。这样HCR就会被启动，产生超长的双链DNA。HCR后，HCR产物的双链不能吸附在GO表面上。因此，HCR 产物发出强烈的荧光信号，该信号取决于 VEGF 165的浓度。通过使用VEGF 165作为模型分析物，该测定提供了一种高度灵敏的VEGF 165荧光检测方法，检测限低至20 pg mL -1。因此可以实现所提出的基于靶标触发的HCR扩增的适体传感策略。成功应用于人血清、尿液和唾液中VEGF 165的测定。因此，它很容易在重大疾病的诊断中得到广泛的应用。
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