Application of riboprinting for the identification of isolates of cutaneousLeishmaniaspp.

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Abstract
Riboprinting is one of several molecular methods that can generate comparative data independently of the complexity of the organism's morphology. Restriction fragment length polymorphism (RFLP) profiles derived from digestion of polymerase chain reaction (PCR) products of the ribosomal 18S fromLeishmaniaspp. yields a typical ‘riboprint’ profile that can vary intraspecifically. A selection of 76 stocks ofL. majorandL. tropica, isolated from patients with cutaneous leishmaniasis, was analysed by riboprinting to assess divergence within and between species.L. majorandL. tropicacould be easily differentiated from each other. Analysis of PCR–RFLP profiles indicated that stocks ofLeishmaniaspp. could be broadly partitioned into 2 species corresponding toL. majorandL. tropica. To test if ribosomal 18S sequences were homogeneous within each species, several isolates of each of theLeishmaniaspp. were digested. Interpretation of the riboprint profiles of the 18S independently amplified by PCR, there would appear to be one restriction pattern present within eachLeishmaniaspp. Homogeneity within copies of the ribosomal 18S within a single genome has, therefore, been demonstrated. The species designation established by riboprinting results were in agreement with the zymodeme analysis of the same isolates. The restriction patterns produced were simple, reproducible and easy to interpret.