Abstract
Conventional methods for the identification of species of Leishmania parasite causing infections have limitations. By using a DNA-based alternative, the present study tries to develop a new tool for this purpose. Thirty-three patients living in Marseilles (in the south of France) were suffering from visceral or cutaneous leishmaniasis. DNA of the parasite in clinical samples (bone marrow, peripheral blood, or skin) from these patients were amplified by PCR and were directly sequenced. The sequences observed were compared to these of 30 strains of the genus causing Old World leishmaniasis collected in Europe, Africa, or Asia. In the analysis of the sequences of the strains, two different sequence patterns for Leishmania infantum, one sequence for Leishmania donovani, one sequence for Leishmania major, two sequences for Leishmania tropica, and one sequence for Leishmania aethiopica were obtained. Four sequences were observed among the strains from the patients: one was similar to the sequence for the L. major strains, two were identical to the sequences for the L. infantum strains, and the last sequence was not observed within the strains but had a high degree of homology with the sequences of the L. infantum and L. donovani strains. The L. infantum strains from all immunocompetent patients had the same sequence. The L. infantum strains from immunodeficient patients suffering from visceral leishmaniasis had three different sequences. This fact might signify that some variants of L. infantum acquire pathogenicity exclusively in immunocompromised patients. To dispense with the sequencing step, a restriction assay with HaeIII was used. Some restriction patterns might support genetic exchanges in members of the genus Leishmania.
Full Text
The Full Text of this article is available as a PDF (369.4 KB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Bastien P., Blaineau C., Taminh M., Rioux J. A., Roizès G., Pagès M. Interclonal variations in molecular karyotype in Leishmania infantum imply a 'mosaic' strain structure. Mol Biochem Parasitol. 1990 Apr;40(1):53–61. doi: 10.1016/0166-6851(90)90079-2. [DOI] [PubMed] [Google Scholar]
- Grimaldi G., Jr, Momen H., Naiff R. D., McMahon-Pratt D., Barrett T. V. Characterization and classification of leishmanial parasites from humans, wild mammals, and sand flies in the Amazon region of Brazil. Am J Trop Med Hyg. 1991 Jun;44(6):645–661. doi: 10.4269/ajtmh.1991.44.645. [DOI] [PubMed] [Google Scholar]
- Guevara P., Alonso G., da Silveira J. F., de Mello M., Scorza J. V., Añez N., Ramírez J. L. Identification of new world Leishmania using ribosomal gene spacer probes. Mol Biochem Parasitol. 1992 Nov;56(1):15–26. doi: 10.1016/0166-6851(92)90150-i. [DOI] [PubMed] [Google Scholar]
- Guizani I., Van Eys G. J., Ismail R. B., Dellagi K. Use of recombinant DNA probes for species identification of Old World Leishmania isolates. Am J Trop Med Hyg. 1994 May;50(5):632–640. doi: 10.4269/ajtmh.1994.50.632. [DOI] [PubMed] [Google Scholar]
- Hassan M. Q., Ghosh A., Ghosh S. S., Gupta M., Basu D., Mallik K. K., Adhya S. Enzymatic amplification of mini-exon-derived RNA gene spacers of Leishmania donovani: primers and probes for DNA diagnosis. Parasitology. 1993 Dec;107(Pt 5):509–517. doi: 10.1017/s0031182000068086. [DOI] [PubMed] [Google Scholar]
- Howard M. K., Ogunkolade W., Bryceson A. D., Davidson R. N., Moody A. H., Miles M. A. A DNA probe for human visceral leishmaniasis. Trans R Soc Trop Med Hyg. 1992 Jan-Feb;86(1):35–36. doi: 10.1016/0035-9203(92)90430-k. [DOI] [PubMed] [Google Scholar]
- Lanotte G., Rioux J. A. Fusion cellulaire chez les Leishmania (Kinetoplastida, Trypanosomatidae). C R Acad Sci III. 1990;310(7):285–288. [PubMed] [Google Scholar]
- Le Blancq S. M., Belehu A., Peters W. Leishmania in the Old World: 3. The distribution of L. aethiopica zymodemes. Trans R Soc Trop Med Hyg. 1986;80(3):360–366. doi: 10.1016/0035-9203(86)90318-4. [DOI] [PubMed] [Google Scholar]
- Le Blancq S. M., Peters W. Leishmania in the Old World: 2. Heterogeneity among L. tropica zymodemes. Trans R Soc Trop Med Hyg. 1986;80(1):113–119. doi: 10.1016/0035-9203(86)90208-7. [DOI] [PubMed] [Google Scholar]
- Le Blancq S. M., Peters W. Leishmania in the Old World: 4. The distribution of L. donovani sensu lato zymodemes. Trans R Soc Trop Med Hyg. 1986;80(3):367–377. doi: 10.1016/0035-9203(86)90320-2. [DOI] [PubMed] [Google Scholar]
- Le Blancq S. M., Schnur L. F., Peters W. Leishmania in the Old World: 1. The geographical and hostal distribution of L. major zymodemes. Trans R Soc Trop Med Hyg. 1986;80(1):99–112. doi: 10.1016/0035-9203(86)90206-3. [DOI] [PubMed] [Google Scholar]
- Lopez M., Inga R., Cangalaya M., Echevarria J., Llanos-Cuentas A., Orrego C., Arevalo J. Diagnosis of Leishmania using the polymerase chain reaction: a simplified procedure for field work. Am J Trop Med Hyg. 1993 Sep;49(3):348–356. doi: 10.4269/ajtmh.1993.49.348. [DOI] [PubMed] [Google Scholar]
- Piarroux R., Azaiez R., Lossi A. M., Reynier P., Muscatelli F., Gambarelli F., Fontes M., Dumon H., Quilici M. Isolation and characterization of a repetitive DNA sequence from Leishmania infantum: development of a visceral leishmaniasis polymerase chain reaction. Am J Trop Med Hyg. 1993 Sep;49(3):364–369. doi: 10.4269/ajtmh.1993.49.364. [DOI] [PubMed] [Google Scholar]
- Piarroux R., Fontes M., Perasso R., Gambarelli F., Joblet C., Dumon H., Quilici M. Phylogenetic relationships between Old World Leishmania strains revealed by analysis of a repetitive DNA sequence. Mol Biochem Parasitol. 1995 Jul;73(1-2):249–252. doi: 10.1016/0166-6851(95)00097-k. [DOI] [PubMed] [Google Scholar]
- Piarroux R., Gambarelli F., Dumon H., Fontes M., Dunan S., Mary C., Toga B., Quilici M. Comparison of PCR with direct examination of bone marrow aspiration, myeloculture, and serology for diagnosis of visceral Leishmaniasis in immunocompromised patients. J Clin Microbiol. 1994 Mar;32(3):746–749. doi: 10.1128/jcm.32.3.746-749.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Pratlong F., Dedet J. P., Marty P., Portús M., Deniau M., Dereure J., Abranches P., Reynes J., Martini A., Lefebvre M. Leishmania-human immunodeficiency virus coinfection in the Mediterranean basin: isoenzymatic characterization of 100 isolates of the Leishmania infantum complex. J Infect Dis. 1995 Jul;172(1):323–326. doi: 10.1093/infdis/172.1.323. [DOI] [PubMed] [Google Scholar]
- Rioux J. A., Lanotte G., Maazoun R., Pasteur N. L'électrophorèse des enzymes dans le genre Leishmania Ross, 1903. Parassitologia. 1985 Jan-Aug;27(1-2):141–156. [PubMed] [Google Scholar]
- Rioux J. A., Lanotte G., Serres E., Pratlong F., Bastien P., Perieres J. Taxonomy of Leishmania. Use of isoenzymes. Suggestions for a new classification. Ann Parasitol Hum Comp. 1990;65(3):111–125. doi: 10.1051/parasite/1990653111. [DOI] [PubMed] [Google Scholar]
- Rodgers M. R., Popper S. J., Wirth D. F. Amplification of kinetoplast DNA as a tool in the detection and diagnosis of Leishmania. Exp Parasitol. 1990 Oct;71(3):267–275. doi: 10.1016/0014-4894(90)90031-7. [DOI] [PubMed] [Google Scholar]
- Smyth A. J., Ghosh A., Hassan M. Q., Basu D., De Bruijn M. H., Adhya S., Mallik K. K., Barker D. C. Rapid and sensitive detection of Leishmania kinetoplast DNA from spleen and blood samples of kala-azar patients. Parasitology. 1992 Oct;105(Pt 2):183–192. doi: 10.1017/s0031182000074096. [DOI] [PubMed] [Google Scholar]
- Spithill T. W., Grumont R. J. Identification of species, strains and clones of Leishmania by characterization of kinetoplast DNA minicircles. Mol Biochem Parasitol. 1984 Jun;12(2):217–236. doi: 10.1016/0166-6851(84)90137-3. [DOI] [PubMed] [Google Scholar]
- Uliana S. R., Affonso M. H., Camargo E. P., Floeter-Winter L. M. Leishmania: genus identification based on a specific sequence of the 18S ribosomal RNA sequence. Exp Parasitol. 1991 Feb;72(2):157–163. doi: 10.1016/0014-4894(91)90133-h. [DOI] [PubMed] [Google Scholar]
- van Eys G. J., Schoone G. J., Kroon N. C., Ebeling S. B. Sequence analysis of small subunit ribosomal RNA genes and its use for detection and identification of Leishmania parasites. Mol Biochem Parasitol. 1992 Mar;51(1):133–142. doi: 10.1016/0166-6851(92)90208-2. [DOI] [PubMed] [Google Scholar]