Abstract
The progression of Pneumocystis carinii pneumonia was temporally monitored and quantified by real-time polymerase chain reaction of P. carinii–specific DNA in oral swabs and lung homogenates from infected rats. DNA levels correlated with the number of P. carinii organisms in the rats’ lungs, as enumerated by microscopic methods. This report is the first of a noninvasive, antemortem method that can be used to monitor infection in a host over time.
Highlights
Pneumocystis pneumonia remains a leading opportunistic infection associated with AIDS patients, even in the era of highly active antiretroviral therapy [1]
The amount of P. carinii–specific DNA quantified by real-time polymerase chain reaction (PCR) in the lung homogenate (LH) samples increased substantially from 0 to 7 weeks, with similar levels after 7 and 10 weeks of immunosuppression (Figure, B)
Progression of Pneumocystis carinii pneumonia measured by enumeration of organisms and real-time PCR of DNA extracted from lung homogenates and oral swabs
Summary
The progression of Pneumocystis carinii pneumonia was temporally monitored and quantified by real-time polymerase chain reaction of P. carinii–specific DNA in oral swabs and lung homogenates from infected rats. We applied the oral swab technique in combination with quantification of organism-specific DNA using real-time polymerase chain reaction (PCR) to monitor the progression of infection in the rat model. The exponential amplification and efficiency of the reactions were determined by evaluating the slope of the curve generated by plotting the log of known concentrations of template DNA vs their CTs [7]. The amount of P. carinii–specific DNA quantified by real-time PCR in the LH samples increased substantially from 0 to 7 weeks, with similar levels after 7 and 10 weeks of immunosuppression (Figure, B). Progression of Pneumocystis carinii pneumonia measured by enumeration of organisms and real-time PCR of DNA extracted from lung homogenates and oral swabs.
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