Document Detail


PCR-based detection in a micro-fabricated platform.
MedLine Citation:
PMID:  18584089     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
We present a novel, on-chip system for the electrokinetic capture of bacterial cells and their identification using the polymerase chain reaction (PCR). The system comprises a glass-silicon platform with a set of micro-channels, -chambers, and -electrodes. A platinum thin film resistor, placed in the proximity of the chambers, is used for temperature monitoring. The whole chip assembly is mounted on a Printed Circuit Board (PCB) and wire-bonded to it. The PCB has an embedded heater that is utilized for PCR thermal cycle and is controlled by a Lab-View program. Similar to our previous work, one set of electrodes on the chip inside the bigger chamber (0.6 microl volume) is used for diverting bacterial cells from a flowing stream into to a smaller chamber (0.4 nl volume). A second set of interdigitated electrodes (in smaller chamber) is used to actively trap and concentrate the bacterial cells using dielectrophoresis (DEP). In the presence of the DEP force, with the cells still entrapped in the micro-chamber, PCR mix is injected into the chamber. Subsequently, PCR amplification with SYBR Green detection is used for genetic identification of Listeria monocytogenes V7 cells. The increase in fluorescence is recorded with a photomultiplier tube module mounted over an epifluorescence microscope. This integrated micro-system is capable of genetic amplification and identification of as few as 60 cells of L. monocytogenes V7 in less than 90 min, in 600 nl volume collected from a sample of 10(4) cfu ml(-1). Specificity trials using various concentrations of L. monocytogenes V7, Listeria innocua F4248, and Escherichia coli O157:H7 were carried out successfully using two different primer sets specific for a regulatory gene of L. monocytogenes, prfA and 16S rRNA primer specific for the Listeria spp., and no cross-reactivity was observed.
Authors:
Shantanu Bhattacharya; Shuaib Salamat; Dallas Morisette; Padmapriya Banada; Demir Akin; Yi-Shao Liu; Arun K Bhunia; Michael Ladisch; Rashid Bashir
Related Documents :
24800179 - The transmembrane channel-like protein family and human papillomaviruses: insights into...
17499489 - A novel, compact disk-like centrifugal microfluidics system for cell lysis and sample h...
18008309 - Microfluidic cell disruption system employing a magnetically actuated diaphragm.
20221559 - Integrated ac electrokinetic cell separation in a closed-loop device.
20875729 - Electric cell-substrate impedance sensing (ecis) based real-time measurement of titer d...
12565999 - Single-cell electroporation.
940549 - Thymidine kinase, dna synthesis and cancer.
2580919 - Adsorption of poly ri:rc on cell membrane participating and nonparticipating in interfe...
19285339 - Glycoprobes as a tool for the study of lectins expressed on tumor cells.
Publication Detail:
Type:  Journal Article     Date:  2008-05-23
Journal Detail:
Title:  Lab on a chip     Volume:  8     ISSN:  1473-0197     ISO Abbreviation:  Lab Chip     Publication Date:  2008 Jul 
Date Detail:
Created Date:  2008-06-27     Completed Date:  2008-09-12     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  101128948     Medline TA:  Lab Chip     Country:  England    
Other Details:
Languages:  eng     Pagination:  1130-6     Citation Subset:  IM    
Affiliation:
Birck Nanotechnology Center, Purdue University, West Lafayette, IN, USA.
Export Citation:
APA/MLA Format     Download EndNote     Download BibTex
MeSH Terms
Descriptor/Qualifier:
Calibration
Cell Separation / instrumentation
Electric Conductivity
Glass / chemistry
Listeria monocytogenes / cytology,  genetics*,  isolation & purification*
Microchip Analytical Procedures / instrumentation*
Microelectrodes
Polymerase Chain Reaction / instrumentation*
Sensitivity and Specificity
Silicon / chemistry
Temperature
Chemical
Reg. No./Substance:
7440-21-3/Silicon

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


Previous Document:  A practical guide to the staggered herringbone mixer.
Next Document:  Rapid and variable-volume sample loading in sieving electrophoresis microchips using negative pressu...