Microfluidic electrochemical assay for rapid detection and quantification of Escherichia coli
Microfluidic electrochemical biosensor for performing Loop-mediated isothermal amplification (LAMP) was developed for the detection and quantification of Escherichia coli. The electrochemical detection for detecting the DNA amplification was achieved using Hoechst 33258 redox molecule and linear swe...
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| Veröffentlicht in: | Biosensors & bioelectronics 2012-01, Vol.31 (1), p.523-528 |
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| creator | Safavieh, Mohammadali Ahmed, Minhaz Uddin Tolba, Mona Zourob, Mohammed |
| description | Microfluidic electrochemical biosensor for performing Loop-mediated isothermal amplification (LAMP) was developed for the detection and quantification of Escherichia coli. The electrochemical detection for detecting the DNA amplification was achieved using Hoechst 33258 redox molecule and linear sweep voltametry (LSV). The DNA aggregation and minor groove binding with redox molecule cause a significant drop in the anodic oxidation of LSV. Unlike other electrochemical techniques, this method does not require the probe immobilization and the detection of the bacteria can be accomplished in a single chamber without DNA extraction and purification steps. The isothermal amplification time has a major role in the quantification of the bacteria. We have shown that we could detect and quantify 24CFU/ml of bacteria and 8.6fg/μl DNA in 60min and 48CFU/ml of bacteria in 35min in LB media and urine samples. We believe that this microfluidic chip has great potential to be used as a point of care diagnostic (POC) device in the clinical/hospital application. |
| doi_str_mv | 10.1016/j.bios.2011.11.032 |
| format | Article |
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The electrochemical detection for detecting the DNA amplification was achieved using Hoechst 33258 redox molecule and linear sweep voltametry (LSV). The DNA aggregation and minor groove binding with redox molecule cause a significant drop in the anodic oxidation of LSV. Unlike other electrochemical techniques, this method does not require the probe immobilization and the detection of the bacteria can be accomplished in a single chamber without DNA extraction and purification steps. The isothermal amplification time has a major role in the quantification of the bacteria. We have shown that we could detect and quantify 24CFU/ml of bacteria and 8.6fg/μl DNA in 60min and 48CFU/ml of bacteria in 35min in LB media and urine samples. 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| subjects | Amplification Bacteria Bacterial Load - instrumentation Biological and medical sciences Biosensing Techniques - instrumentation Biosensor Biosensors Biotechnology Conductometry - instrumentation Electrochemical detection Equipment Design Equipment Failure Analysis Escherichia coli Escherichia coli - isolation & purification Fundamental and applied biological sciences. Psychology Methods. Procedures. Technologies Microfluidics Reproducibility of Results Sensitivity and Specificity Various methods and equipments |
| title | Microfluidic electrochemical assay for rapid detection and quantification of Escherichia coli |
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