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用于环境传输研究的 DNA 标记的 PLGA 颗粒的制备、检测和分析。

Fabrication, detection, and analysis of DNA-labeled PLGA particles for environmental transport studies.

机构信息

Department of Land, Air, and Water Resources, University of California, Davis, Davis, CA 95616, USA.

Department of Land, Air, and Water Resources, University of California, Davis, Davis, CA 95616, USA.

出版信息

J Colloid Interface Sci. 2018 Sep 15;526:207-219. doi: 10.1016/j.jcis.2018.04.059. Epub 2018 Apr 18.

Abstract

Poly(lactic-co-glycolic acid) (PLGA) particle carriers of synthetic DNA have recently received increased attention for environmental applications due to their biodegradability, customizability, and nearly limitless number of uniquely identifiable "labels". In this paper, we present methodologies for the preparation of DNA-labeled particles, control of particle size, extraction of DNA-labels, and analysis via quantitative polymerase chain reaction (qPCR). Characterization and analysis of the DNA-labeled particles reveal spherical particles of diameters ranging from 60 to 1000 nm, with consistent zeta potentials around -45 mV, that are stable to aggregation, even in the presence of concentrated mono- and divalent cations. A highly correlated and consistent relationship between particle concentration and DNA-label count was observed, with a detection range spanning 7 orders of magnitude, from 0.01 to 10,000 mg/L (10-10 particles/μL). The results of two environmental applications of the DNA-labeled particles are also presented, highlighting their feasibility for use in environmental studies. Whether exploring size-dependent transport phenomena or identifying potential pathogen transport pathways, the DNA-labeled particle approach presented here provides a powerful tool for the identification of overlapping particle signals at a range of concentrations.

摘要

聚(乳酸-共-乙醇酸)(PLGA)颗粒载体作为合成 DNA 的载体,由于其生物降解性、可定制性以及几乎无限数量的独特可识别“标签”,最近在环境应用中受到了越来越多的关注。在本文中,我们提出了制备 DNA 标记颗粒、控制颗粒大小、提取 DNA 标记物以及通过定量聚合酶链反应(qPCR)进行分析的方法。对 DNA 标记颗粒的表征和分析表明,这些颗粒为直径在 60 至 1000nm 之间的球形颗粒,其表面zeta 电位约为-45mV,具有抗聚集的稳定性,即使在存在浓度较高的单价和二价阳离子的情况下也是如此。观察到颗粒浓度与 DNA 标记计数之间存在高度相关且一致的关系,检测范围跨越 7 个数量级,从 0.01 至 10,000mg/L(10-10 个颗粒/μL)。本文还介绍了 DNA 标记颗粒在两个环境应用中的结果,突出了其在环境研究中的应用可行性。无论是探索尺寸依赖性的输运现象还是识别潜在病原体的输运途径,本文提出的 DNA 标记颗粒方法都为在一系列浓度下识别重叠的颗粒信号提供了有力的工具。

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