Molecular detection of atypical bacteria and viruses linked to community-acquired pneumonia

Abstract

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.