EMPLOYMENT OF THE POLYMERASE CHAIN-REACTION IN THE MOLECULAR DIFFERENTIATION OF REPTILIAN HEMOGREGARINES AND ITS APPLICATION TO PREVENTATIVE ZOOLOGICAL MEDICINE

This study involved the development and evaluation of a polymerase cha in reaction (PCR)-based strategy for typing morphologically similar sp ecies and isolates of reptilian hemogregarines. This typing is importa nt to zoological medicine because hemogregarines can be transmitted ac ross reptile species and can produce clinically significant disease in unnatural host species. Several hemogregarine isolates obtained from different reptilian host species were molecularly characterized at the ribosomal DNA (rDNA) locus by PCR amplification and restriction enzym e analysis. Amplification of the rDNA from five different hemogregarin e isolates resulted in the robust production of 590-bp DNA fragments. Restriction enzyme digestion and electrophoretic analysis of the ampli fied 590-bp fragments resulted in banding patterns specific for hemogr egarines obtained from different host species. The fact that hemogrega rines can be molecularly distinguished at the rDNA locus is significan t. A PCR-based ribotyping assay system could be employed as a surveill ance device for detecting interspecific transmission among reptile pop ulations.