Berkeley Sensor and Actuator Center, Department of Bioengineering, Department of Electrical Engineering and Computer Science, University of California, Berkeley, California 94720, United States.
ACS Nano. 2012 Sep 25;6(9):7607-14. doi: 10.1021/nn203833d. Epub 2012 Aug 21.
Early cancer diagnosis is very important for the prevention or mitigation of metastasis. However, effective and efficient methods are needed to improve the diagnosis and assessment of cancer. Here, we report a single-step detection method using a nanoplasmonic aptamer sensor (aptasensor), targeting a vascular endothelial growth factor-165 (VEGF(165)), a predominant biomarker of cancer angiogenesis. Our single-step detection is accomplished by (1) specific target recognition by an aptamer-target molecule interaction and (2) direct readouts of the target recognition. The readout is achieved by inactivation of surface plasmon enhancement of fluorescent probes preattached to the aptamers. Our aptasensor provides the appropriate sensitivity for clinical diagnostics with a wide range of linear detection from 25 pg/mL to 25 μg/mL (=from 1.25 pM to 1.25 μM), high specificity for VEGF(165) against PDGF-BB, osteopontin (OPN), VEGF(121), NaCl, and temporal/thermal/biological stability. In experiments with 100% serum and saliva from clinical samples, readouts of the aptasensor and an ELISA for VEGF(165) show good agreement within the limit of the ELISA kit. We envision that our developed aptasensor holds utilities for point-of-care cancer prognostics by incorporating simplicity in detection, low-cost for test, and required small sample volumes.
早期癌症诊断对于预防或减轻转移非常重要。然而,需要有效的方法来提高癌症的诊断和评估。在这里,我们报告了一种使用纳米等离子体适体传感器(aptasensor)的单步检测方法,该传感器靶向血管内皮生长因子-165(VEGF(165)),这是癌症血管生成的主要生物标志物。我们的单步检测是通过(1)适体与靶分子相互作用的特异性靶识别和(2)靶识别的直接读出来完成的。读出是通过预先附着在适体上的荧光探针的表面等离子体增强的失活来实现的。我们的适体传感器提供了适当的灵敏度,用于临床诊断,具有从 25 pg/mL 到 25 μg/mL(即从 1.25 pM 到 1.25 μM)的宽线性检测范围,对 PDGF-BB、骨桥蛋白(OPN)、VEGF(121)、NaCl 和时间/热/生物稳定性具有高特异性。在使用来自临床样本的 100%血清和唾液的实验中,aptasensor 的读出值和 ELISA 检测 VEGF(165)的值在 ELISA 试剂盒的限制内很好地一致。我们设想,我们开发的适体传感器通过检测的简单性、测试的低成本和所需的小样本量,在即时癌症预后方面具有实用性。