A PCR SCHEME FOR DIFFERENTIATION OF ORGANISMS BELONGING TO THE MYCOPLASMA-MYCOIDES CLUSTER

Rapid and specific detection of organisms belonging to the Mycoplasma mycoides cluster, among them Mycoplasma (M.) mycoides subsp. mycoides SC, the agent of contagious bovine pleuropneumonia, is an important co ndition for effective control of the respective animal diseases. In an effort to improve diagnosis, a PCR identification scheme for these my coplasmas was developed. A set of primer combinations derived from the CAP-21 genomic regions of member organisms of the Mycoplasma mycoides cluster was selected by means of which complete differentiation withi n the cluster can be accomplished. Nested PCR involving cluster-specif ic amplification at the first stage and group-specific amplification u sing internal primers at the second stage was shown to be applicable f or identification of all six groups forming the cluster. For example, external primers P1/P2 and internal primers P6/P7 were used to disting uish M. mycoides subsp. mycoides SC from LC strains, Using the present PCR procedure, identification of mycoplasmas could be considerably ac celerated in comparison to conventional methods (5 h vs. one week) and specificity was also improved.