DETECTION OF MYCOPLASMA CONTAMINATIONS BY THE POLYMERASE CHAIN-REACTION

The polymerase chain reaction (PCR) has been used for the general dete ction of Mollicutes. 25 Mycoplasma and Acholeplasma species were detec ted including important contaminants of cell cultures such as M, orale , M. arginini, M. hyorhinis, M. fermentans, A. laidlawii and additiona l human and animal mycoplasmas. PCR reactions were performed using a s et of nested primers defined from conserved regions of the 16S rRNA ge ne. The detection limit was determined to be I fg mycoplasma DNA, whic h is equivalent to 1-2 genome copies of the 16S rRNA coding region. Th e identity of the amplification products was confirmed by agarose gel electrophoresis and restriction enzyme analysis. DNA from closely and distantly related micro-organisms did not give rise to specific amplif ication products. The method presented here offers a much more sensiti ve, specific and rapid assay for the detection of mycoplasmas than the existing ones.