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Background   Mycoplasma pneumoniae  and  Chlamydophila pneumoniae  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  M. pneumoniae  and/or C.  pneumoniae  DNA in clinical respiratory samples.    Objective  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.    Methods  A total of 146 clinical respiratory specimens, collected from 1997 to 2007, suspected of  C. pneumoniae  or  M. pneumoniae  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  C. pneumoniae  and culture as the reference method for the analysis of  M. pneumoniae . For discordant analysis specimens were re-tested using another commercial multiplex PCR, the PneumoBacter-1 assay (Seegene, Korea).    Results  Following discordant analysis, the sensitivity of the ProPneumo-1 assay for pathogens,  C. pneumoniae  or  M. pneumoniae , was 100%. The specificity of the ProPneumo-1 assay, however, was 100% for  C. pneumoniae  and 98% for  M. pneumoniae . The limits of detection were 1 genome equivalent (Geq) per reaction for pathogens,  M. pneumoniae  and  C. pneumoniae . Due to the multipex format of the ProPneumo-1 assay, we identified 5 additional positive specimens, 2  C. pneumoniae  in the  M. pneumoniae -negative pool and 3  M. pneumoniae  in the  C. pneumoniae -negative pool.    Conclusion  The ProPneumo-1 assay is a rapid, sensitive and effective method for the simultaneous detection of  M. pneumoniae  and  C. pneumoniae  directly in respiratory specimens.

annals of clinical microbiology and antimicrobials

Verification of the ProPneumo-1 assay for the simultaneous detection of Mycoplasma pneumoniae and Chlamydophila pneumoniae in clinical respiratory specimens