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UBC Theses and Dissertations

Analytical approaches for the management of resistant cytomegalovirus infection Au, Eugene


Cytomegalovirus is a member of the herpesviridae family which is prevalent in the human population. Following primary viral infection, cytomegalovirus persists in a latent state, from which the virus can reactivate resulting in renewed shedding. While cytomegalovirus rarely causes disease in the immunocompetent population, infection in newborns and immunocompromised patients can result in significant morbidity and mortality. Current strategies for the management of cytomegalovirus infection utilise drugs such as ganciclovir which inhibit viral DNA polymerase. As with other viruses, incomplete suppression of replication during long-term use of antiviral drugs favours the development of resistance resulting in drug failure. In the management of resistant cytomegalovirus infection, the determination of adequate drug delivery warrants attention, and thus, a sensitive and selective capillary electrophoresis assay for ganciclovir concentration in plasma was developed and validated. The assay utilises solid phase extraction and detection by ultraviolet absorbance, and has a lower limit of quantification of 40 ng/ml. Along with pharmacokinetic monitoring, another consideration in the management of drug resistance is the identification of viral susceptibility, and quantitative polymerase chain reaction was used to investigate the effects of ganciclovir on cytomegalovirus DNA. For this investigation, molecular cloning was conducted to produce plasmid-virus DNA, which was used as a quantitative standard in assessing the inhibition of cytomegalovirus DNA replication by ganciclovir in reference and resistant virus strains. In addition to the determination of susceptibility, quantitative polymerase chain reaction can also be applied in the screening of novel antiviral agents against cytomegalovirus, which are required due to continued concerns surrounding antiviral resistance.

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