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从血液中通过介电泳分离 DNA 和纳米颗粒。

Dielectrophoretic isolation of DNA and nanoparticles from blood.

机构信息

Department of Bioengineering, University of California San Diego, La Jolla, CA 92093-0448, USA.

出版信息

Electrophoresis. 2012 Aug;33(16):2482-90. doi: 10.1002/elps.201100700.

Abstract

The ability to effectively detect disease-related DNA biomarkers and drug delivery nanoparticles directly in blood is a major challenge for viable diagnostics and therapy monitoring. A DEP method has been developed which allows the rapid isolation, concentration and detection of DNA and nanoparticles directly from human and rat whole blood. Using a microarray device operating at 20 V peak-to-peak and 10 kHz, a wide range of high molecular weight (HMW)-DNA and nanoparticles were concentrated into high-field regions by positive DEP, while the blood cells were concentrated into the low-field regions by negative DEP. A simple fluidic wash removes the blood cells while the DNA and nanoparticles remain concentrated in the DEP high-field regions where they can be detected by fluorescence. HMW-DNA could be detected at 260 ng/mL, which is a detection level suitable for analysis of disease-related cell-free circulating DNA biomarkers. Fluorescent 40 nm nanoparticles could be detected at 9.5 × 10(9) particles/mL, which is a level suitable for monitoring drug delivery nanoparticles. The ability to rapidly isolate and detect DNA biomarkers and nanoparticles from undiluted whole blood will benefit many diagnostic applications by significantly reducing sample preparation time and complexity.

摘要

有效地检测血液中与疾病相关的 DNA 生物标志物和药物输送纳米粒子是可行的诊断和治疗监测的主要挑战。已经开发出一种 DEP 方法,可以直接从人和大鼠全血中快速分离、浓缩和检测 DNA 和纳米粒子。使用工作在 20 V 峰峰值和 10 kHz 的微阵列装置,可以通过正 DEP 将高分子量(HMW)-DNA 和纳米粒子浓缩到高场区,而血细胞则通过负 DEP 浓缩到低场区。简单的流体冲洗可去除血细胞,而 DNA 和纳米粒子则保留在 DEP 高场区,通过荧光进行检测。可以检测到 260 ng/mL 的 HMW-DNA,这是适合分析与疾病相关的无细胞循环 DNA 生物标志物的检测水平。可以检测到 9.5×10(9)个/mL 的荧光 40nm 纳米粒子,这是适合监测药物输送纳米粒子的检测水平。从未稀释的全血中快速分离和检测 DNA 生物标志物和纳米粒子的能力将通过大大减少样品制备时间和复杂性,使许多诊断应用受益。

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