A NEW APPROACH FOR THE IDENTIFICATION AND THE DIAGNOSIS OF EIMERIA-MEDIA PARASITE OF THE RABBIT

In this work, we described a new approach for the isolation of a speci es-specific probe for the Eimeria media parasite of the rabbit based o n the use of the random amplified polymorphic DNA (RAPD) technique. A specific fragment of 800 bp of the studied species was isolated after RAPD and then cloned and DIG-radiolabeled. After dot-blotting, we obse rved that this probe was specific for E. media. Sequencing of the 3' a nd 5' ends of this probe enabled the determination of two primers thar could be used in a PCR reaction. The amplified product of 750 bp was specific for E. media. The use of these primers and of our probe allow ed the detection of a very small number of oocysts. With a new protoco l of DNA purification, 10 purified oocysts were detected by PCR. The e fficiency of the amplification was not changed when two species were m ixed. The threshold of detection of oocysts in fecal matter was equal to 30. (C) 1996 Academic Press, Inc.