Development of a novel approach multiplex late-pcr electrochemical-enzyme based dna sensor for sequence specific detection of vibrio cholerae

Yean, Chan Yean (2015) Development of a novel approach multiplex late-pcr electrochemical-enzyme based dna sensor for sequence specific detection of vibrio cholerae. Project Report. Pusat Pengajian Kesihatan. (Submitted)

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Abstract

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

Item Type: Monograph (Project Report)
Uncontrolled Keywords: vibrio cholerae
Subjects: R Medicine
R Medicine > RC Internal medicine
Divisions: Kampus Kesihatan (Health Campus) > Pusat Pengajian Sains Kesihatan (School of Health Sciences) > Monograph
Depositing User: Mr Husnan Budin
Date Deposited: 31 Mar 2024 02:15
Last Modified: 31 Mar 2024 02:15
URI: http://eprints.usm.my/id/eprint/59940

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