Establishment of TaqMan-MGB real-time PCR assay to detect Mycoplasma pneumoniae

Objective To establish a sensitive and specific TaqMan-MGB real-time PCR assay to detect Mycoplasma pneumoniae from clinical specimens. Methods By analyzing the RepMP23 gene sequence of M. pneumoniae isolates, an optimized real-time PCR assay was designed. The specificity, sensitivity and amplification efficiency of this assay were evaluated with standard concentration M. pneumoniae DNA, and compared with routine TaqMan real-time PCR assay. Results The detection limitation of this TaqMan-MGB real-time PCR assay was about 0.2 cfu, about 10 times higher than routine TaqMan real-time PCR assay. The specificity of the assay seemed to be 100% in the detection of 23 common respiratory tract pathogens and human DNA. Conclusion The TaqMan-MGB real-time PCR assay was superior than TaqMan real-time PCR method. It is suitable for the detection of M. pneumoniae in clinical specimens.