循环肿瘤细胞
单细胞测序
计算生物学
单细胞分析
多重位移放大
微流控
全血
微流控芯片
全基因组测序
生物
基因组
基因
外显子组测序
突变
癌症
DNA提取
遗传学
细胞
聚合酶链反应
材料科学
纳米技术
免疫学
转移
作者
Li Ren,Fu‐Jun Jia,Weikai Zhang,Fan Shi,Zhiguo Fang,Hong Zhao,Zhiyuan Hu,Zewen Wei
出处
期刊:Lab on a Chip
[The Royal Society of Chemistry]
日期:2019-01-01
卷期号:19 (19): 3168-3178
被引量:24
摘要
Whole-genome sequencing on circulating tumor cells (CTCs) at the single cell level has recently been found helpful for precision medicine, as the oncogenic profiles of single CTCs are useful for discovering oncogenic mutation heterogeneities and guiding/adjusting cancer treatment. To overcome the limits of existing methods of single CTC sequencing, in which CTC enrichment, identification and gene amplification are performed by discrete modules, this study presents a novel method in which all processing steps from blood sample collection to preparation of gene amplification products for sequencers are finished in a single microfluidic chip. This microfluidic chip comprehensively performs blood filtering, CTC enrichment, CTC identification/isolation, CTC lysis and whole genome amplification (WGA) at the single cell level. By sequencing single CTCs from clinical blood samples with pointing key driver and drug-resistance mutations, the novel microfluidic chip was validated to be capable of genetically profiling single CTCs with minimum cell loss/human labor, and more importantly, high accuracy and repeatability, which are crucial factors for promoting clinical application of single CTC sequencing.
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