Amplification of Mycoplasma iowae using polymerase chain reaction.

Based on sequence data of Mycoplasma iowae recombinant DNA probe, pMI-12, two 25-base primers were synthesized for use in the polymerase chain reaction (PCR). An M. iowae species-specific 299-base pair product was amplified by the primers. An annealing temperature of 58.5 C was critical for detecting all members of this heterogeneous species while maintaining specificity of the M. iowae PCR. The minimum amount of target DNA detected by M. iowae PCR was 1 pg, which was 1000 times more sensitive than the dot-blot assay using M. iowae recombinant DNA probes.