Development of a novel approach multiplex late-pcr electrochemical-enzyme based dna sensor for sequence specific detection of vibrio cholerae
Phase 1: Development of a Multiplex Linear-After-The-Exponential (LATE)- Polymerase Chain Reaction (PCR) • To design specific primers and optimize a multiplex LATE-PCR which is capable of simultaneously detecting the presence of V. cholerae, as well as cholera toxin and internal control genes....
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| Main Author: | |
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| Format: | Monograph |
| Language: | English |
| Published: |
Pusat Pengajian Kesihatan
2015
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| Subjects: | |
| Online Access: | http://eprints.usm.my/59940/1/DR%20CHAN%20YEAN%20YEAN%20-%20e.pdf http://eprints.usm.my/59940/ |
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| Summary: | Phase 1: Development of a Multiplex Linear-After-The-Exponential (LATE)- Polymerase Chain
Reaction (PCR)
• To design specific primers and optimize a multiplex LATE-PCR which is capable of
simultaneously detecting the presence of V. cholerae, as well as cholera toxin and internal control
genes.
Phase 2: Development of a Mixed Self-Assembled Monolayer (SAM) on Disposable Screen-Printed
Gold Electrode (SPGE)
• To design thiol-modified capture probe for sequence-specific hybridization with target LATEPCR
amplicons.
• To optimize the immobilization of capture probe DNAs on gold electrode surface using Self-
Assembled Monolayer (SAM) system.
Phase 3: Development of a Multiplex Electrochemical-Enzyme based DNA sensor
• To optimize the hybridization of labeled LATE-PCR amplicons to the capture probe DNAs.
• To optimize the simultaneous detection of multiple redox reactions catalyzed by alkaline
phosphatase and horseradish peroxidase by amperometry.
Phase 4: Evaluation of the Multiplex Electrochemical-Enzyme based DNA sensor
• To perform analytical evaluation of the multiplex electrochemical DNA hybridization
genosensor.
• To evaluate the performance of the multiplex electrochemical DNA hybridization genosensor
using clinical samples.
Objective Achieved
(Please state the extent to which the project objectives were achieved)
Phase 1: Development of a Multiplex Linear-After-The-Exponential (LATE)- Polymerase Chain
Reaction (PCR)
• To design specific primers and optimize a multiplex LATE-PCR which is capable of
simultaneously detecting the presence of V. cholerae, as well as cholera toxin and internal control
genes -100% achieved as per planned
Phase 2: Development of a Mixed Self-Assembled Monolayer (SAM) on Disposable Screen-Printed
Gold Electrode (SPGE)
• To design thiol-modified capture probe for sequence-specific hybridization with target LATEPCR
amplicons.-100% achieved as per planned
• To optimize the immobilization of capture probe DNAs on gold electrode surface using Self-
Assembled Monolayer (SAM) system -100% achieved as per planned
Phase 3: Development of a Multiplex Electrochemical-Enzyme based DNA sensor
• To optimize the hybridization of labeled LATE-PCR amplicons to the capture probe DNAs -
100% achieved as per planned
• To optimize the simultaneous detection of multiple redox reactions catalyzed by alkaline
phosphatase and horseradish peroxidase by amperometry -100% achieved as per planned
Phase 4: Evaluation of the Multiplex Electrochemical-Enzyme based DNA sensor
• To perform analytical evaluation of the multiplex electrochemical DNA hybridization
genosensor -100% achieved as per planned
• To evaluate the performance of the multiplex electrochemical DNA hybridization genosensor
using clinical samples -100% achieved as per planned |
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