Journal
WORLD JOURNAL OF MICROBIOLOGY & BIOTECHNOLOGY
Volume 28, Issue 4, Pages 1699-1706Publisher
SPRINGER
DOI: 10.1007/s11274-011-0978-x
Keywords
Vibrio cholerae; Enzyme-based electrochemical genosensor; Probe-based sandwich-type hybridization; Asymmetric PCR amplicon; Carbon screen-printed electrode; Chronoamperometry
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Funding
- Universiti Sains Malaysia (USM), Malaysia [1001/PPSP/813020, 1001/PPSP/8144004]
- Institute of Postgraduate Studies (IPS), USM
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A disposable horseradish peroxidase (HRP)-based electrochemical genosensor was developed for chronoamperometric detection of single-stranded asymmetric lolB gene PCR amplicon (118 bp in length) of the food-borne pathogen, Vibrio cholerae. A two-step sandwich-type hybridization strategy using two specific probes was employed for specific detection of the target single-stranded DNA (ssDNA). The analytical performances of the detection platform have been evaluated using a synthetic ssDNA (ST3) which was identical to the target single-stranded amplicon and a total of 19 bacterial strains. Under optimal condition, ST3 was calibrated with a dynamic range of 0.4883-15.6250 nM. By coupling asymmetric PCR amplification, the probe-based electrochemical genosensor was highly specific to the target organism (100% specificity) and able to detect as little as 0.85 ng/mu l of V. cholerae genomic DNA.
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