Isolation and Detection of Mycoplasma pneumoniae from Cell Culture by Culture and PCR

Mycoplasma pneumoniae is one of the major species of mycoplasmas which could contaminate cell cultures. Identification of
M. pneuminae is significant because Mycoplasmas could contaminate and have unsuitable effect on the cell cultures. Rapid detection of these contaminations is so important and it could be significant role in preventing and controlling of contamination in cell cultures. The aim of this study is isolation and detection of
M .pneumoniae by culture and PCR. 82 cell cultures collected and cultured in PPLO broth media supplemented for M. pnemoniae isolation.

The bacteria DNAs were extracted by phenol/chloroform method and the PCR assay amplifying the conserved region of 16S rRNA gene was applied for the detection of Mycoplasma genus in 163bp fragment and M. pnemoniae in 543bp fragment by using of MPF and MPR primers which obtained from P1 adhesin gen.

Of the 82 samples 48(58.5%) yielded one of the potentially pathogenic Mycoplasmas evaluated for using Mycoplasma genus PCR as diagnostic method, and from 48 samples which were positive in genus of Mycoplasma, M. pneumoniae did not detect  from those sample by using those primers and there is not any M. pnemoniae detected in this study.

Also results of this study indicate that PCR could be an alternative method instead of the culture because according to the results of this study, PCR has high accuracy and speed for detecting M. pneumoniae.