Rapid Detection of Urinary Tract Infections Using Isotachophoresis and Molecular Beacons

被引:82
作者
Bercovici, M. [1 ,2 ]
Kaigala, G. V. [1 ,2 ]
Mach, K. E. [2 ]
Han, C. M. [1 ]
Liao, J. C. [2 ]
Santiago, J. G. [1 ]
机构
[1] Stanford Univ, Dept Mech Engn, Stanford, CA 94305 USA
[2] Stanford Univ, Dept Urol, Stanford, CA 94305 USA
基金
美国国家科学基金会;
关键词
CAPILLARY ISOTACHOPHORESIS; DNA; ACID; HYBRIDIZATION; OPTIMIZATION; KINETICS; MATRICES; CHLORIDE;
D O I
10.1021/ac200253x
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
We present a novel assay for rapid detection and identification of bacterial urinary tract infections using isotachophoresis (ITP) and molecular beacons. We applied on-chip ITP to extract and focus 16S rRNA directly from bacterial lysate and used molecular beacons to achieve detection of bacteria specific sequences. We demonstrated detection of E. coli in bacteria cultures as well as in patient urine samples in the clinically relevant range 1E6-1E8 cfu/mL. For bacterial cultures we further demonstrate quantification in this range. The assay requires minimal sample preparation (a single centrifugation and dilution), and can be completed, from beginning of lysing to detection, in under 15 min. We believe that the principles presented here can be used for design of other rapid diagnostics or detection methods for pathogenic diseases.
引用
收藏
页码:4110 / 4117
页数:8
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