Highly sensitive and quantitative detection of rare pathogens through agarose droplet microfluidic emulsion PCR at the single-cell level

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	Highly sensitive and quantitative detection of rare pathogens through agarose droplet microfluidic emulsion PCR at the single-cell level
	Zhu, Zhi ; Zhang, Wenhua ; Leng, Xuefei ; Zhang, Mingxia ; Guan, Zhichao ; Lu, Jiangquan ; Yang, Chaoyong James ; Zhu Z(朱志) ; Yang CY(杨朝勇)
刊名	http://dx.doi.org/10.1039/c2lc40461c
	2012
关键词	POLYMERASE-CHAIN-REACTION ESCHERICHIA-COLI O157-H7 DNA AMPLIFICATION MOLECULE PCR PARALLEL WATER EFFICIENT GENOME
英文摘要	Natural Science Foundation of China [21075104]; National Basic Research Program of China [2010CB732402]; Natural Science Foundation of Fujian Province for Distinguished Young Scholars [2010 J06004]; National Found for Fostering Talents of Basic Science [J1030415]; Genetic alternations can serve as highly specific biomarkers to distinguish fatal bacteria or cancer cells from their normal counterparts. However, these mutations normally exist in very rare amount in the presence of a large excess of non-mutated analogs. Taking the notorious pathogen E. coli O157:H7 as the target analyte, we have developed an agarose droplet-based microfluidic ePCR method for highly sensitive, specific and quantitative detection of rare pathogens in the high background of normal bacteria. Massively parallel singleplex and multiplex PCR at the single-cell level in agarose droplets have been successfully established. Moreover, we challenged the system with rare pathogen detection and realized the sensitive and quantitative analysis of a single E. coli O157:H7 cell in the high background of 100 000 excess normal K12 cells. For the first time, we demonstrated rare pathogen detection through agarose droplet microfluidic ePCR. Such a multiplex single-cell agarose droplet amplification method enables ultra-high throughput and multi-parameter genetic analysis of large population of cells at the single-cell level to uncover the stochastic variations in biological systems.
语种	英语
出版者	LAB CHIP
内容类型	期刊论文
源URL	[http://dspace.xmu.edu.cn/handle/2288/88369]
专题	化学化工－已发表论文
推荐引用方式 GB/T 7714	Zhu, Zhi,Zhang, Wenhua,Leng, Xuefei,et al. Highly sensitive and quantitative detection of rare pathogens through agarose droplet microfluidic emulsion PCR at the single-cell level[J]. http://dx.doi.org/10.1039/c2lc40461c,2012.
APA	Zhu, Zhi.,Zhang, Wenhua.,Leng, Xuefei.,Zhang, Mingxia.,Guan, Zhichao.,...&杨朝勇.(2012).Highly sensitive and quantitative detection of rare pathogens through agarose droplet microfluidic emulsion PCR at the single-cell level.http://dx.doi.org/10.1039/c2lc40461c.
MLA	Zhu, Zhi,et al."Highly sensitive and quantitative detection of rare pathogens through agarose droplet microfluidic emulsion PCR at the single-cell level".http://dx.doi.org/10.1039/c2lc40461c (2012).