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Heparin-Bead Extraction Enhanced Fluorogenic Aptamer–Thrombin Composite Reporter Enables Sensitive and Rapid Detection of Functional Antithrombin
ACS Sensors ( IF 8.2 ) Pub Date : 2024-08-23 , DOI: 10.1021/acssensors.4c01294 He Yan 1 , Zimu Tian 1 , Emma Ziegenbein 2 , Ying Liu 1 , Amanda Santana 1 , Adam S Veige 1 , Bruce Spiess 3 , Weihong Tan 1 , Yong Zeng 1, 2, 4
ACS Sensors ( IF 8.2 ) Pub Date : 2024-08-23 , DOI: 10.1021/acssensors.4c01294 He Yan 1 , Zimu Tian 1 , Emma Ziegenbein 2 , Ying Liu 1 , Amanda Santana 1 , Adam S Veige 1 , Bruce Spiess 3 , Weihong Tan 1 , Yong Zeng 1, 2, 4
Affiliation
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Antithrombin (AT) deficiency in the extracorporeal circulation during cardiac surgery leads to uncontrolled inflammation and vascular damage in patients. AT levels decrease in sepsis, major trauma, extracorporeal membrane oxygenation, and eclampsia. Monitoring plasma AT levels facilitates the accurate restoration of AT to baseline values through precise supplementation. Traditional methods of chromogenic assay and enzyme-linked immunosorbent assay (ELISA) kits encounter challenges, such as interference, inconsistency, and delayed response times, making real-time, reliable antithrombin monitoring a clinical gap. To address this critical need, we develop a heparin-bead extraction enhanced fluoroGenic aptamer–thrombin composite reporter (HExGATOR) for the rapid, sensitive, and precise detection of functional AT in plasma. Our design employs thrombin-binding aptamers and a fluorescence “turn on” technology such that a signal is produced upon the interaction of AT with the otherwise “turned off” aptamer–thrombin complex. The prominent signal-background interference originating from plasma is remarkably diminished by using a heparin-bead solid-phase extraction of AT. We achieved highly sensitive and rapid detection of AT in 5 to 20 min with a limit of detection of 15.11 nM. This approach offers a promising alternative to traditional AT tests in clinical settings, potentially facilitating personalized anticoagulant therapy.
中文翻译：
肝素微珠提取增强型荧光适配体-凝血酶复合报告基因可实现灵敏、快速的功能性抗凝血酶检测
心脏手术期间体外循环中的抗凝血酶 （AT） 缺乏会导致患者不受控制的炎症和血管损伤。脓毒症、严重创伤、体外膜肺氧合和子痫的 AT 水平降低。监测血浆 AT 水平有助于通过精确补充将 AT 准确恢复到基线值。显色测定和酶联免疫吸附测定 （ELISA） 试剂盒的传统方法遇到了干扰、不一致和响应时间延迟等挑战，这使得实时、可靠的抗凝血酶监测成为临床空白。为了满足这一关键需求，我们开发了一种肝素珠提取增强型荧光显配子-凝血酶复合报告基因 （HExGATOR），用于快速、灵敏和精确地检测血浆中的功能性 AT。我们的设计采用凝血酶结合适配体和荧光“开启”技术，使得在 AT 与原本“关闭”的适配体-凝血酶复合物相互作用时产生信号。使用 AT 的肝素珠固相萃取可显著减少源自血浆的突出信号背景干扰。我们在 5 至 20 分钟内实现了 AT 的高灵敏度和快速检测，检测限为 15.11 nM。这种方法为临床环境中的传统 AT 测试提供了一种有前途的替代方案，可能促进个性化的抗凝治疗。
更新日期：2024-08-23
中文翻译：
肝素微珠提取增强型荧光适配体-凝血酶复合报告基因可实现灵敏、快速的功能性抗凝血酶检测
心脏手术期间体外循环中的抗凝血酶 （AT） 缺乏会导致患者不受控制的炎症和血管损伤。脓毒症、严重创伤、体外膜肺氧合和子痫的 AT 水平降低。监测血浆 AT 水平有助于通过精确补充将 AT 准确恢复到基线值。显色测定和酶联免疫吸附测定 （ELISA） 试剂盒的传统方法遇到了干扰、不一致和响应时间延迟等挑战，这使得实时、可靠的抗凝血酶监测成为临床空白。为了满足这一关键需求，我们开发了一种肝素珠提取增强型荧光显配子-凝血酶复合报告基因 （HExGATOR），用于快速、灵敏和精确地检测血浆中的功能性 AT。我们的设计采用凝血酶结合适配体和荧光“开启”技术，使得在 AT 与原本“关闭”的适配体-凝血酶复合物相互作用时产生信号。使用 AT 的肝素珠固相萃取可显著减少源自血浆的突出信号背景干扰。我们在 5 至 20 分钟内实现了 AT 的高灵敏度和快速检测，检测限为 15.11 nM。这种方法为临床环境中的传统 AT 测试提供了一种有前途的替代方案，可能促进个性化的抗凝治疗。
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