Multi-center evaluation of one commercial and 12 in-house real-time PCR assays for detection of Mycoplasma pneumoniae

•We compared 13 real-time PCR assays to detect M. pneumoniae.•Participating laboratories used identical DNA samples and a standardized protocol.•All assays were able to detect at least 20 genomes per reaction.

Detection of Mycoplasma pneumoniae by real-time PCR is not yet standardized across laboratories. We have implemented a standardization protocol to compare the performance of thirteen commercial and in-house approaches. Despite differences on threshold values of samples, all assays were able to detect at least 20 M. pneumoniae genomes per reaction.