Amplification of the 16S-23S Spacer Region in rRNA Operons of Mycoplasmas by the Polymerase Chain Reaction

Nucleotide sequences of the spacer region between 16S and 23S DNA in the ribosomal RNA operons of mycoplasmas were identified by analysis of products of the polymerase chain reaction (PCR) amplified from the corresponding regions of 12 species of this family. Three common PCR primers, F1, F2, and R1, were designed by analysis of similarity between these sequences. Primers F1 and R1 produced fragments of 340 to 660 bp when the DNA of each species was used as the template. Specific amplification of the spacer region was confirmed by a second round of PCR in which the amplified products were used as the templates and F2 and R1 were used as the primers. No discrete band was observed in electrophoresis when human or mouse DNA served as the template with use of primers F1 and R2, which suggests that many mycoplasmal species that sometimes contaminate a culture of eukaryotic cells can be detected by the PCR.