| Summary: | <p>Abstract</p> <p>Background</p> <p><it>Mycoplasma pneumoniae </it>and <it>Chlamydophila pneumoniae </it>are major causes of lower and upper respiratory infections that are difficult to diagnose using conventional methods such as culture. The ProPneumo-1 (Prodesse, Waukesha, WI) assay is a commercial multiplex real-time PCR assay for the simultaneous detection of <it>M. pneumoniae </it>and/or C. <it>pneumoniae </it>DNA in clinical respiratory samples.</p> <p>Objective</p> <p>The aim of this study was to evaluate the sensitivity and specificity of the ProPneumo-1, a newly developed commercial multiplex real-time PCR assay.</p> <p>Methods</p> <p>A total of 146 clinical respiratory specimens, collected from 1997 to 2007, suspected of <it>C. pneumoniae </it>or <it>M. pneumoniae </it>infections were tested retrospectively. Nucleic acid was extracted using an automated NucliSense easyMag (bioMerieux, Netherlands). We used a "Home-brew" monoplex real-time assay as the reference method for the analysis of <it>C. pneumoniae </it>and culture as the reference method for the analysis of <it>M. pneumoniae</it>. For discordant analysis specimens were re-tested using another commercial multiplex PCR, the PneumoBacter-1 assay (Seegene, Korea).</p> <p>Results</p> <p>Following discordant analysis, the sensitivity of the ProPneumo-1 assay for pathogens, <it>C. pneumoniae </it>or <it>M. pneumoniae</it>, was 100%. The specificity of the ProPneumo-1 assay, however, was 100% for <it>C. pneumoniae </it>and 98% for <it>M. pneumoniae</it>. The limits of detection were 1 genome equivalent (Geq) per reaction for pathogens, <it>M. pneumoniae </it>and <it>C. pneumoniae</it>. Due to the multipex format of the ProPneumo-1 assay, we identified 5 additional positive specimens, 2 <it>C. pneumoniae </it>in the <it>M. pneumoniae</it>-negative pool and 3 <it>M. pneumoniae </it>in the <it>C. pneumoniae</it>-negative pool.</p> <p>Conclusion</p> <p>The ProPneumo-1 assay is a rapid, sensitive and effective method for the simultaneous detection of <it>M. pneumoniae </it>and <it>C. pneumoniae </it>directly in respiratory specimens.</p>
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