Development Of A Glass Fibre Membrane-Based Lateral Flow Biosensor For The Detection Of Abelled PCR Amplicons Using Vibrio Cholerae As The Model Organism
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Date
2010-12
Authors
Chua, Ang Lim
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Journal ISSN
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Publisher
Universiti Sains Malaysia
Abstract
In recent years, molecular techniques have played an increasing role in diagnosis of infection diseases and polymerase chain reaction (PCR) is one of the major techniques used. However major setbacks such as requiring trained personel, cold-chain storage and cumbersome detection protocols hampered the general use of PCR in place of conventional methods. PCR products are commonly detected using agarose gel eletrophoresis methods involving carcinogenic sthidium bromide and hsrmful ultraviolet illumination. Hence, the objective of this study was to develope and evaluate a themostabilised PCR assay and a lateral flow-based device for detection of amplicons. Vibro cholerae was used as a model organism to develope a molecular detection kit named the cholerae PCR-ICT-DNA assay. A multiplex PCR with an incorperated internal amplification control (IC) and utilised hapen modified primers was developed and optimised. The PCR was thermostabilised and evaluated for analytical specificity, a latral flow device was developed and optimised using FUSION 5 glass fibre membrane (Whatman, UK).
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Keywords
An insreasing role in diagnosis of infection diseases , and polymerase chain reaction (PCR)