The MOE Key Laboratory of Spectrochemical Analysis & Instrumentation, the Key Laboratory of Chemical Biology of Fujian Province, State Key Laboratory of Physical Chemistry of Solid Surfaces, Collaborative Innovation Center of Chemistry for Energy Materials, Department of Chemical Biology, Department of Chemical Engineering, College of Chemistry and Chemical Engineering, Pen-Tung Sah Institute of Micro-Nano Science and Technology , Xiamen University , Xiamen 361005 , China.
Institute of Molecular Medicine, Renji Hospital , Shanghai Jiao Tong University School of Medicine , Shanghai 200127 , China.
Anal Chem. 2019 May 7;91(9):6003-6011. doi: 10.1021/acs.analchem.9b00470. Epub 2019 Apr 8.
As the third-generation nucleic acid amplification technology, digital polymerase chain reaction (PCR) has been widely adopted in the analysis of nucleic acids. However, further application of this powerful technology is hindered due to the limitation of surfactants. Here, for the first time, we propose the use of colloidosomes self-assembled from fluorinated silica nanoparticle for digital PCR to address this limitation. A one-step fluorinated silica nanoparticle synthesis method is proposed, which is much more convenient and reproducible compared with the synthesis of conventional fluorine-based surfactants. Fluorinated silica nanoparticles facilitate the formation of colloidosomes with excellent stability capable of enduring the rapid temperature changes associated with the PCR and avoiding material exchange (cross-talk) between droplets for high-fidelity analysis. The colloidosome digital PCR method was developed using these colloidosomes as highly parallel reactors for single-copy nucleic acid amplification and rare mutant detection. The method is robust and accurate, and it offers possibilities for a great variety of applications, such as gene expression studies, single cell analysis, and circulating tumor DNA detection.
作为第三代核酸扩增技术，数字聚合酶链反应（PCR）已广泛应用于核酸分析。然而，由于表面活性剂的限制，这项强大技术的进一步应用受到了阻碍。在这里，我们首次提出使用氟硅纳米粒子自组装的胶体囊泡来解决这一限制。提出了一种一步法氟硅纳米粒子合成方法，与传统含氟表面活性剂的合成方法相比，该方法更加方便且重现性更好。氟硅纳米粒子有利于形成具有极好稳定性的胶体囊泡，能够耐受与 PCR 相关的快速温度变化，并避免液滴之间的物质交换（串扰），从而实现高保真度分析。使用这些胶体囊泡作为高度并行的反应器，我们开发了胶体囊泡数字 PCR 方法，用于单拷贝核酸扩增和稀有突变检测。该方法稳健且准确，为各种应用提供了可能性，例如基因表达研究、单细胞分析和循环肿瘤 DNA 检测。
