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Application of an arbitrarily-primed polymerase chain reaction to mycoplasma identification and typing within the
G Rawadi1, B Lemercier, D Roulland-Dussoix
1Roussel Uclaf, Domaine thérapeutique Immunologie, Romainville, France.
Abstract:
An arbitrarily-primed polymerase chain reaction (AP-PCR) was developed using a primer pair, Mlip1 and Mlip4, for members of the Mycoplasma mycoides cluster, a group containing important pathogens of small and large ruminants. Parameters that influence the reproducibility of this assay were optimized: magnesium, primer and template concentrations, and pH. AP-PCR fingerprinting, carried out on a number of strains of each of the six species or subspecies belonging to the mycoides cluster, allowed the typing of strains within each group. The AP-PCR assay showed that the cluster can be divided into two groups: (i) high and (ii) no genomic polymorphism variation. In addition, specific polymorphic bands for members of species or subspecies included in this cluster were amplified by this AP-PCR method, thus allowing their identification.