| Summary: | M.Sc.(Med.), Faculty of Health Sciences, University of the Witwatersrand, 2009 === Community-acquired pneumonia (CAP) is a major cause of morbidity and mortality worldwide.
Knowledge of the predominant agents associated with CAP locally is essential, as it represents the
basis for empiric antibiotic treatment. The objective of this study was to establish polymerase
chain reaction (PCR)-based methods that could be used to identify CAP pathogens. Real-time
PCR assays were developed to detect 10 viral and 5 non-viral pathogens as well as 2 internal
controls using SYBR Green I and TaqMan probes, in singleplex and multiplex reactions. Six
multiplex assays, with sensitivities of 1-10 copies/μl, were successfully developed to
simultaneously detect 12 organisms. These reactions were used to test a limited number of patient
and simulated samples. Data from the real-time PCR methods compared favourably to those from
commercially available conventional PCR kits. These detection methods could be used to
complement each other in prevalence studies and in selected diagnostic applications.
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