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Application of riboprinting for the identification of isolates of cutaneous Leishmania spp.

Published online by Cambridge University Press:  09 October 2003

L. F. NIMRI  and
H. D. F. H. SCHALLIG
L. F. NIMRI
Affiliation:
Department of Medical Laboratory Sciences, Jordan University of Science and Technology, P.O. Box 3030, Irbid 22110, Jordan
H. D. F. H. SCHALLIG
Affiliation:
Royal Tropical Institute (KIT), Department of Biomedical Research, Meibergdreef 39, 1105 AZ Amsterdam, The Netherlands
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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 from Leishmania spp. yields a typical ‘riboprint’ profile that can vary intraspecifically. A selection of 76 stocks of L. major and L. tropica, isolated from patients with cutaneous leishmaniasis, was analysed by riboprinting to assess divergence within and between species. L. major and L. tropica could be easily differentiated from each other. Analysis of PCR–RFLP profiles indicated that stocks of Leishmania spp. could be broadly partitioned into 2 species corresponding to L. major and L. tropica. To test if ribosomal 18S sequences were homogeneous within each species, several isolates of each of the Leishmania spp. were digested. Interpretation of the riboprint profiles of the 18S independently amplified by PCR, there would appear to be one restriction pattern present within each Leishmania spp. 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.

Keywords

Leishmania majorLeishmania tropicapolymerase chain reactionrestriction enzymesrestriction fragment length polymorphismriboprinting
Type
Research Article
Information
Parasitology , Volume 127 , Issue 3 , September 2003 , pp. 201 - 205
Copyright
© 2003 Cambridge University Press

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