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Title: Development of DNA microarray system for the diagnosis of virus infections of the central nervous system
Author: Abdel-Hakeem, Mohammed Abdel-Razek
ISNI:       0000 0004 2747 0846
Awarding Body: University of Nottingham
Current Institution: University of Nottingham
Date of Award: 2010
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The increasing numbers of neurovirulent viruses, their similar clinical manifestations and the limited number of detectable viruses per assay increase the urge for a more comprehensive and rapid diagnostic assay. This will be reflected on patient management and community savings. Although PCR is a powerful tool for nucleic acid amplification, its specific amplification confines its use to a small number of viruses. Whole genome amplification using random techniques may represent suitable comprehensive alternatives to PCR when combined with the specific DNA microarray technologies. In this project, variable parameters in the field of DNA microarray technology were investigated such as probe design, surface chemistry and hybridisation optimisation. Target preparation in terms of random amplification and labelling was also investigated. The possibility of moving the assay from the expensive fluorescent protocols to cheaper alternatives more suited to widespread use was explored using manual colorimetric assays in place of automated fluorescent detection procedures. After extensive developmental work, a single protocol was proved to differentiate between herpes simplex virus 1, herpes simplex 2, Epstein Barr virus, cytomegalovirus, varicella zoster virus, human herpesvirus 6, and coxsackievirus. The sensitivity of the assay was close to the sensitivity of reference laboratories using PCR based techniques. The colorimetric assay also provided a cost-effective sensitive format that is capable of differentiating between various viruses. This assay offers an opportunity to develop a single process capable of simultaneous detection of a large number of possible central nervous viruses in a shorter period of time. The initial colorimetric results render the assay more applicable in laboratories with low technical facilities.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available