Proximal Bacterial Lysis and Detection in Nanoliter Wells Using Electrochemistry

被引:33
作者
Besant, Justin D. [1 ]
Das, Jagotamoy [2 ]
Sargent, Edward H. [3 ]
Kelley, Shana O. [1 ,2 ,4 ]
机构
[1] Univ Toronto, Inst Biomat & Biomed Engn, Toronto, ON M5S 3M2, Canada
[2] Univ Toronto, Dept Pharmaceut Sci, Leslie Dan Fac Pharm, Toronto, ON M5S 3M2, Canada
[3] Univ Toronto, Dept Elect & Comp Engn, Fac Appl Sci & Engn, Toronto, ON M5S 3M2, Canada
[4] Univ Toronto, Dept Biochem, Fac Med, Toronto, ON M5S 3M2, Canada
基金
加拿大自然科学与工程研究理事会;
关键词
bacterial detection; cell lysis; electrochemical detection; microelectrodes; PATHOGENIC DNA; CULTURE; POINT; CARE; NANOPARTICLES; PERFORMANCE; DIAGNOSIS; SYSTEM;
D O I
10.1021/nn4035298
中图分类号
O6 [化学];
学科分类号
0703 ;
摘要
Rapid and direct genetic analysis of low numbers of bacteria using chip-based sensors is limited by the slow diffusion of mRNA molecules. Long incubation times are required in dilute solutions in order to collect a sufficient number of molecules at the sensor surface to generate a detectable signal. To overcome this barrier here we present an integrated device that leverages electrochemistry-driven lysis less than 50 mu m away from electrochemical nucleic acid sensors to overcome this barrier. Released intracellular mRNA can diffuse the short distance to the sensors within minutes, enabling rapid and sensitive detection. We validate this strategy through direct lysis and detection of E. coli mRNA at concentrations as low as 0.4 CFU/mu L in 2 min, a clinically relevant combination of speed and sensitivity for a sample-to-answer molecular analysis approach.
引用
收藏
页码:8183 / 8189
页数:7
相关论文
共 25 条
  • [1] The rpoB gene as a tool for clinical microbiologists
    Adekambi, Toidi
    Drancourt, Michel
    Raoult, Didier
    [J]. TRENDS IN MICROBIOLOGY, 2009, 17 (01) : 37 - 45
  • [2] Anker J. N. J., 2008, NAT MATER, V7, P8
  • [3] Dissociation and Degradation of Thiol-Modified DNA on Gold Nanoparticles in Aqueous and Organic Solvents
    Bhatt, Nishi
    Huang, Po-Jung Jimmy
    Dave, Neeshma
    Liu, Juewen
    [J]. LANGMUIR, 2011, 27 (10) : 6132 - 6137
  • [4] A multicenter trial to compare blood culture with polymerase chain reaction in severe human sepsis
    Bloos, Frank
    Hinder, Frank
    Becker, Karsten
    Sachse, Svea
    Dessap, Armand Mekontso
    Straube, Eberhard
    Cattoir, Vincent
    Brun-Buisson, Christian
    Reinhart, Konrad
    Peters, Georg
    Bauer, Michael
    [J]. INTENSIVE CARE MEDICINE, 2010, 36 (02) : 241 - 247
  • [5] Nanostructured biomolecular detectors: pushing performance at the nanoscale
    Cederquist, Kristin B.
    Kelley, Shana O.
    [J]. CURRENT OPINION IN CHEMICAL BIOLOGY, 2012, 16 (3-4) : 415 - 421
  • [6] Chung HJ, 2013, NAT NANOTECHNOL, V8, P369, DOI [10.1038/nnano.2013.70, 10.1038/NNANO.2013.70]
  • [7] Electrochemical DNA sensors
    Drummond, TG
    Hill, MG
    Barton, JK
    [J]. NATURE BIOTECHNOLOGY, 2003, 21 (10) : 1192 - 1199
  • [8] Eckert DJ, 2010, EXPERT REV MOL DIAGN, V10, P399, DOI [10.1586/erm.10.24, 10.1586/ERM.10.24]
  • [9] Genetic Analysis of H1N1 Influenza Virus from Throat Swab Samples in a Microfluidic System for Point-of-Care Diagnostics
    Ferguson, B. Scott
    Buchsbaum, Steven F.
    Wu, Ting-Ting
    Hsieh, Kuangwen
    Xiao, Yi
    Sun, Ren
    Soh, H. Tom
    [J]. JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 2011, 133 (23) : 9129 - 9135
  • [10] Microfluidic designs and techniques using lab-on-a-chip devices for pathogen detection for point-of-care diagnostics
    Foudeh, Amir M.
    Didar, Tohid Fatanat
    Veres, Teodor
    Tabrizian, Maryam
    [J]. LAB ON A CHIP, 2012, 12 (18) : 3249 - 3266