A surface topography assisted droplet manipulation platform for biomarker detection and pathogen identification

被引:0
作者
Zhang, Yi
Park, Seungkyung [2 ]
Liu, Kelvin
Tsuan, Jennifer
Yang, Samuel [2 ]
Wang, Tza-Huei [1 ]
机构
[1] Johns Hopkins Univ, Dept Mech Engn, Dept Biomed Engn,Ctr Canc Nanotechnol Excellence, Sidney Kimmel Comprehens Canc Ctr, Baltimore, MD 21218 USA
[2] Johns Hopkins Univ, Dept Emergency Med, Baltimore, MD USA
基金
美国国家科学基金会;
关键词
TOTAL ANALYSIS SYSTEMS; ON-A-CHIP; PERSONALIZED MEDICINE; MICROFLUIDIC SYSTEMS; DNA AMPLIFICATION; PCR; LAB; TECHNOLOGIES; PURIFICATION; DEVICE;
D O I
10.1039/c0lc00296h
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
This paper reports a droplet microfluidic, sample-to-answer platform for the detection of disease biomarkers and infectious pathogens using crude biosamples. The platform exploited the dual functionality of silica superparamagnetic particles (SSP) for solid phase extraction of DNA and magnetic actuation. This enabled the integration of sample preparation and genetic analysis within discrete droplets, including the steps of cell lysis, DNA binding, washing, elution, amplification and detection. The microfluidic device was self contained, with all reagents stored in droplets, thereby eliminating the need for fluidic coupling to external reagent reservoirs. The device incorporated unique surface topographic features to assist droplet manipulation. Pairs of micro-elevations were created to form slits that facilitated efficient splitting of SSP from droplets. In addition, a compact sample handling stage, which integrated the magnet manipulator, the droplet microfluidic device and a Peltier thermal cycler, was built for convenient droplet manipulation and real-time detection. The feasibility of the platform was demonstrated by analysing ovarian cancer biomarker Rsf-1 and detecting Escherichia coli with real time polymerase chain reaction and real time helicase dependent amplification.
引用
收藏
页码:398 / 406
页数:9
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