Detection of Mycoplasma meleagridis by polymerase chain reaction

Abstract

A pair of 32 base primers was synthesized based on the DNA sequence data of a Mycoplasma meleagridis (MM) species-specific recombinant, pMM-2. The primers were used in a MM-polymerase chain reaction (PCR) to amplify a target DNA of approximately 850 bp. Annealing temperatures ranging from 58°C to 61°C could be used for the MM-PCR without loss of specifity. The primers amplified 1 ng of DNA from 17 strains of MM, but not 10 ng of DNA from 16 heterologous species of avian mycoplasma, pUC8 plasmid, lambda phage or calf thymus DNA. The minimum amount of target DNA detected by MM-PCR was 10 fg, which indicated that this procedure was 100000 times more sensitive than dot blot methodology using an MM recombinant DNA probe.