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从血液中通过介电泳分离和检测 cfc-DNA 纳米颗粒生物标志物和病毒。

Dielectrophoretic isolation and detection of cfc-DNA nanoparticulate biomarkers and virus from blood.

机构信息

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

出版信息

Electrophoresis. 2013 Apr;34(7):1076-84. doi: 10.1002/elps.201200444.

Abstract

Dielectrophoretic (DEP) microarray devices allow important cellular nanoparticulate biomarkers and virus to be rapidly isolated, concentrated, and detected directly from clinical and biological samples. A variety of submicron nanoparticulate entities including cell free circulating (cfc) DNA, mitochondria, and virus can be isolated into DEP high-field areas on microelectrodes, while blood cells and other micron-size entities become isolated into DEP low-field areas between the microelectrodes. The nanoparticulate entities are held in the DEP high-field areas while cells are washed away along with proteins and other small molecules that are not affected by the DEP electric fields. DEP carried out on 20 μL of whole blood obtained from chronic lymphocytic leukemia patients showed a considerable amount of SYBR Green stained DNA fluorescent material concentrated in the DEP high-field regions. Whole blood obtained from healthy individuals showed little or no fluorescent DNA materials in the DEP high-field regions. Fluorescent T7 bacteriophage virus could be isolated directly from blood samples, and fluorescently stained mitochondria could be isolated from biological buffer samples. Using newer DEP microarray devices, high-molecular-weight DNA could be isolated from serum and detected at levels as low as 8-16 ng/mL.

摘要

介电泳(DEP)微阵列设备可快速从临床和生物样本中直接分离、浓缩和检测重要的细胞纳米颗粒生物标志物和病毒。各种亚微米级纳米颗粒实体,包括无细胞循环(cfc)DNA、线粒体和病毒,都可以在微电极上的 DEP 高场区中分离出来,而血细胞和其他微米大小的实体则在微电极之间的 DEP 低场区中分离出来。纳米颗粒实体被保留在 DEP 高场区中,而细胞则随着不受 DEP 电场影响的蛋白质和其他小分子一起被冲走。对来自慢性淋巴细胞白血病患者的 20 μL 全血进行的 DEP 实验表明,相当数量的 SYBR Green 染色 DNA 荧光物质集中在 DEP 高场区。来自健康个体的全血在 DEP 高场区中显示出很少或没有荧光 DNA 物质。荧光 T7 噬菌体病毒可直接从血液样本中分离出来,荧光染色的线粒体可从生物缓冲液样本中分离出来。使用更新的 DEP 微阵列设备,可以从血清中分离出高分子量 DNA,并以低至 8-16 ng/mL 的水平进行检测。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9af7/4517469/e7f7a41d5d9a/nihms501182f1.jpg

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