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RAPD method useful for distinguishing Leishmania species: design of specific primers for L. braziliensis

Published online by Cambridge University Press:  05 December 2003

E. MARTINEZ ,
V. ALONSO ,
A. QUISPE ,
M. C. THOMAS ,
R. ALONSO ,
J. E. PIÑERO ,
A. C. GONZÁLEZ ,
A. ORTEGA  and
B. VALLADARES
E. MARTINEZ
Affiliation:
Department of Parasitology, Faculty of Pharmacy, University of La Laguna, Avda. Francisco Sánchez s/n, CP38271 La Laguna, Tenerife, Spain
V. ALONSO
Affiliation:
Department of Parasitology, Faculty of Pharmacy, University of La Laguna, Avda. Francisco Sánchez s/n, CP38271 La Laguna, Tenerife, Spain
A. QUISPE
Affiliation:
Department of Parasitology, Faculty of Pharmacy, University of La Laguna, Avda. Francisco Sánchez s/n, CP38271 La Laguna, Tenerife, Spain
M. C. THOMAS
Affiliation:
Unidad Mixta de Investigaciones Médicas. Hospital Clínico Universitario San Cecilio. Dr Oloriz, 16, CP18012 Granada. Spain
R. ALONSO
Affiliation:
Department of Parasitology, Faculty of Pharmacy, University of La Laguna, Avda. Francisco Sánchez s/n, CP38271 La Laguna, Tenerife, Spain
J. E. PIÑERO
Affiliation:
Department of Parasitology, Faculty of Pharmacy, University of La Laguna, Avda. Francisco Sánchez s/n, CP38271 La Laguna, Tenerife, Spain
A. C. GONZÁLEZ
Affiliation:
Department of Parasitology, Faculty of Pharmacy, University of La Laguna, Avda. Francisco Sánchez s/n, CP38271 La Laguna, Tenerife, Spain
A. ORTEGA
Affiliation:
Department of Parasitology, Faculty of Pharmacy, University of La Laguna, Avda. Francisco Sánchez s/n, CP38271 La Laguna, Tenerife, Spain
B. VALLADARES
Affiliation:
Department of Parasitology, Faculty of Pharmacy, University of La Laguna, Avda. Francisco Sánchez s/n, CP38271 La Laguna, Tenerife, Spain
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Abstract

The technique of Random Amplification Polymorphic DNA allows fragments of the genome to be amplified by means of polymerase chain reaction (PCR) without previous knowledge of their sequences. The protozoa of the genus Leishmania present great genetic variability, making it difficult to characterize the different species. A method is developed with a single 10-mers long primer, which allows the species L. braziliensis, L. mexicana, L. infantum, L. tropica, L. chagasi, L. amazonensis and L. major to be differentiated. These products amplified by RAPD have also facilitated the design of some primers that amplify L. braziliensis DNA exclusively.

Keywords

RAPDPCRprimerLeishmania braziliensis
Type
Research Article
Information
Parasitology , Volume 127 , Issue 6 , December 2003 , pp. 513 - 517
Copyright
2003 Cambridge University Press

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References

REFERENCES

ALTSCHUL, S. F., MADDEN, T. L., SCHAFFER, A. A., ZHANG, J., ZHANG, Z., MILLER, W. & LIPMAN, D. J. ( 1997). Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Research 25, 3389–3402.CrossRefGoogle Scholar
BANULS, A. L., HIDE, M. & TIBAYRENC, M. ( 1999). Molecular epidemiology and evolutionary genetics of Leishmania parasites. International Journal for Parasitology 29, 1137–1147.CrossRefGoogle Scholar
DOS SANTOS, J. B., NIENHUIS, J. & SLOCUM, M. K. ( 1994). Comparison of RAPD and RFLP genetic markers in determining genetic similarity among Brassica oleracea L. genotypes. Theoretical and Applied Genetics 87, 909–915.CrossRefGoogle Scholar
EISENBERGER, C. L. & JAFFE, C. L. ( 1999). Leishmania: identification of Old World species using a Permissively Primed Intergenic Polymerase Chain Reaction. Experimental Parasitology 91, 70–77.CrossRefGoogle Scholar
GOLDBERG, S. S. & CHIARI, E. ( 1980). Growth and isolate of single colonies of Trypanosoma cruzi on solid medium. Journal of Parasitology 66, 677–679.CrossRefGoogle Scholar
GONZALEZ, A. C., MARTINEZ, E., CARMELO, E., PIÑERO, J. E., ALONSO, V., DEL CASTILLO, A. & VALLADARES, B. ( 2002). Analysis of NLS and rRNA binding motifs in L25 ribosomal protein from Leishmania (Viannia) braziliensis: investigation of its diagnostic capabilities. Parasitology 125, 51–57.CrossRefGoogle Scholar
HANOFI, R., BARRHOUMI, M., BELHDJ ALI, S. & GUIZANI, I. ( 2001). Molecular analyses of old world Leishmania RAPD markers and development of a PCR assay selective for parasites of the L. donovani species. Experimental Parasitology 98, 90–99.CrossRefGoogle Scholar
HIDE, M., BAÑULS, A. L. & TIBAYRENC, M. ( 2001). Genetic heterogeneity and phylogenetic status of Leishmania (Leishmania) infantum zymodeme MON-1: epidemiological implications. Parasitology 123, 425–432.CrossRefGoogle Scholar
MAURICIO, I. L., STOTHARD, J. R. & MILES, M. A. ( 2000). The strange case of Leishmania chagasi. Parasitology Today 16, 188–189.CrossRefGoogle Scholar
PIÑERO, J. E., MARTINEZ, E., PACHECO, R., ARAGÓN, Z., DE ARMAS, F., DEL CASTILLO, A. & VALLADARES, B. ( 1999). PCR-ELISA for diagnosis of mucocutaneous leishmaniasis. Acta Tropica 25, 21–29.CrossRefGoogle Scholar
RODRIGUES, E. H., FELINTO DE BRITO, M. E., MENDOÇA, M. G., WERKHAUSER, R. P., COUTINHO, E. M., SOUZA, W. V., MILITAO DE ALBUQUERQUE, M. DE F., JARDIM, M. L. & ABATH, F. G. ( 2002). Evaluation of PCR for diagnosis of American cutaneous leishmaniasis in an area of endemicity in northeastern Brazil. Journal of Clinical Microbiology 40, 3572–3576.CrossRefGoogle Scholar
SAMBROOK, J., FRITSCH, E. F. & MANIATIS, T. ( 1989). Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory Press, NY.
VOLPINI, A. C., DE AZEREDO PASSOS, V. M. & ROMANHA, A. J. ( 2001). Attempt to differentiate Leishmania (Leishmania) amazonensis, L. (L.) chagasi, Leishmania (Viannia) braziliensis and L. (V.) guyanensis using the SSR-PCR technique. Parasitology Research 87, 1056–1059.Google Scholar
WILLIAMS, J. G., KUBELIK, A. R., LIVAK, K. J., RAFALSKI, J. A. & TINGEEY, S. V. ( 1990). DNA polymorphins amplified by arbitrary primers are useful as genetic markers. Nucleic Acids Research 18, 6531–6535.CrossRefGoogle Scholar