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Restriction Endonuclease Analysis of Leishmania Kinetoplast DNA Characterizes Parasites Responsible for Visceral and Cutaneous Disease^*

Peter R. Jackson, John A. Wohlhieter, Joan E. Jackson, Perry Sayles, Carter L. Diggs AND Wayne T. Hockmeyer

Departments of Immunology
Bacterial Immunology
Parasitology, Walter Reed Army Institute of Research, Washington, D.C. 20307

The kinetoplast DNA (kDNA) from promastigotes of Leishmania responsible for Old and New World cutaneous and visceral disease was characterized to determine if species and strains causing similar or different diseases could be identified. Restriction enzymes were used to digest kDNA into fragments that were separated into characteristic banding patterns after electrophoresis in agarose or linear gradient polyacrylamide gels. Hybridization was conducted with a ³²P-kDNA probe and kDNA fragments transferred from agarose gels to nitrocellulose paper. Leishmania species causing cutaneous diseases in the New and Old Worlds all had different kDNA digest patterns. Visceralizing Leishmania from the New and Old Worlds also had different kDNA restriction fragment patterns although Leishmania donovani parasites with similar fragment patterns were isolated from several humans from central Kenya. Nucleotide sequences were shared among kDNA networks from L. donovani, Leishmania d. chagasi, Leishmania d. infantum, Leishmania tropica, and Leishmania major as determined by hybridization with a ³²P-kDNA probe from L. donovani. However, no hybridization was detected between the L. donovani ³²P-kDNA probe and kDNA from Leishmania aethiopica or Leishmania braziliensis panamensis. Leishmania characterization results for the same isolates from the published literature were compared and kinetoplast DNA analysis was found to be one of the most sensitive procedures for species and strain identification.

Accepted for publication March 2, 1984.

^* Financial support for portions of this investigation was received from the UNDP/World Bank/WHO Special Programme for Research and Training in Tropical Diseases.

In conducting research described in this report, the investigators adhered to the Guide for Laboratory Animal Facilities and Care as promulgated by the Committee of the Guide for Laboratory Animals and Care of the Institute of Laboratory Animal Resources, National Academy of Sciences, National Research Council.

The views of the authors do not purport to reflect the positions of the Department of the Army or the Department of Defense (Para. 4-3, AR 360-5).

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