Magnetic bead-based DNA detection with multi-layers quantum dots labeling for rapid detection of Escherichia coli O157:H7.

This work demonstrated the feasibility of detecting 250zM Escherichia coli O157:H7 eaeA target DNA by using a magnetic bead-based DNA detection assay with designed labeling strategy within 40-60min. The magnetic beads were used as the solid support for the binding probe and isolated the target DNA from the sample. The detection signals could be amplified from the multi-layers biotin-streptavidin conjugated quantum dots based on binding with specific designed biotinlyted linker. This assay method would provide a simple, rapid, and ultra-sensitive detection method for DNA or other biomolecular analysis.

[1]  Yuzuru Takamura,et al.  Peptide nucleic acid modified magnetic beads for intercalator based electrochemical detection of DNA hybridization , 2004 .

[2]  D. Balding,et al.  HLA Sequence Polymorphism and the Origin of Humans , 2006 .

[3]  J. Lakowicz Principles of fluorescence spectroscopy , 1983 .

[4]  Tza-Huei Wang,et al.  Multiplexed hybridization detection with multicolor colocalization of quantum dot nanoprobes. , 2005, Nano letters.

[5]  R. Tsien,et al.  The Fluorescent Toolbox for Assessing Protein Location and Function , 2006, Science.

[6]  C. Mirkin,et al.  Nanoparticle-Based Bio-Bar Codes for the Ultrasensitive Detection of Proteins , 2003, Science.

[7]  M Wiedmann,et al.  Detection of Escherichia coli O157:H7 by multiplex PCR , 1995, Journal of clinical microbiology.

[8]  A. Bhagwat Simultaneous detection of Escherichia coli O157:H7, Listeria monocytogenes and Salmonella strains by real-time PCR. , 2003, International journal of food microbiology.

[9]  Martin A. M. Gijs,et al.  Magnetic bead handling on-chip: new opportunities for analytical applications , 2004 .

[10]  A. M. Young Taxonomy and Natural History , 2004, Science.

[11]  Mansel W. Griffiths,et al.  Rapid Detection of Escherichia coli O157:H7 with Multiplex Real-Time PCR Assays , 2002, Applied and Environmental Microbiology.

[12]  Yanbin Li,et al.  A self-assembled monolayer-based piezoelectric immunosensor for rapid detection of Escherichia coli O157:H7. , 2004, Biosensors & bioelectronics.

[13]  J. Yu,et al.  Cloning and characterization of the eae gene of enterohaemorrhagic Escherichia coli O157:H7 , 1992, Molecular microbiology.

[14]  Chad A Mirkin,et al.  Bio-bar-code-based DNA detection with PCR-like sensitivity. , 2004, Journal of the American Chemical Society.

[15]  X. Mao,et al.  A nanoparticle amplification based quartz crystal microbalance DNA sensor for detection of Escherichia coli O157:H7. , 2006, Biosensors & bioelectronics.

[16]  Yanbin Li,et al.  Immunobiosensor chips for detection of Escherichia coil O157:H7 using electrochemical impedance spectroscopy. , 2002, Analytical chemistry.

[17]  Jeeseong Hwang,et al.  High-sensitivity bacterial detection using biotin-tagged phage and quantum-dot nanocomplexes. , 2006, Proceedings of the National Academy of Sciences of the United States of America.

[18]  H. Chart,et al.  Bacteria in biology, biotechnology and medicine , 1998 .

[19]  M. Louie,et al.  PCR Detection of Escherichia coli O157:H7 Directly from Stools: Evaluation of Commercial Extraction Methods for Purifying Fecal DNA , 2000, Journal of Clinical Microbiology.

[20]  Hiroshi Ito,et al.  Development of an enzymatic reaction device using magnetic bead-cluster handling , 2006 .

[21]  Gehua Wang,et al.  Detection in Escherichia coli of the Genes Encoding the Major Virulence Factors, the Genes Defining the O157:H7 Serotype, and Components of the Type 2 Shiga Toxin Family by Multiplex PCR , 2002, Journal of Clinical Microbiology.

[22]  Nicole Jaffrezic-Renault,et al.  Biosensors and Bio-Bar Code Assays Based on Biofunctionalized Magnetic Microbeads , 2007, Sensors (Basel, Switzerland).

[23]  P. Fratamico,et al.  Detection of Escherichia coli 0157:H7 using a surface plasmon resonance biosensor , 1998 .

[24]  V. Sharma,et al.  Detection of enterohemorrhagic Escherichia coli O157:H7 by using a multiplex real-time PCR assay for genes encoding intimin and Shiga toxins. , 2003, Veterinary microbiology.

[25]  Paul Singleton,et al.  Bacteria in biology, biotechnology, and medicine , 1995 .

[26]  Shuming Nie,et al.  Single-bead immunoassays using magnetic microparticles and spectral-shifting quantum dots. , 2007, Journal of agricultural and food chemistry.

[27]  Ying Sun,et al.  Design and performances of immunoassay based on SPR biosensor with magnetic microbeads. , 2007, Biosensors & bioelectronics.

[28]  K. Livak,et al.  A PCR-based assay for the detection of Escherichia coli Shiga-like toxin genes in ground beef , 1996, Applied and environmental microbiology.

[29]  C. Yamashiro,et al.  PCR-Based DNA Amplification and Presumptive Detection of Escherichia coli O157:H7 with an Internal Fluorogenic Probe and the 5′ Nuclease (TaqMan) Assay , 1998, Applied and Environmental Microbiology.

[30]  I Karube,et al.  Detection of PCR products of Escherichia coli O157:H7 in human stool samples using surface plasmon resonance (SPR). , 2000, FEMS immunology and medical microbiology.

[31]  S. Nie,et al.  Quantum dot bioconjugates for ultrasensitive nonisotopic detection. , 1998, Science.