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ARE CYTOCHROME B GENE SEQUENCING AND POLYMORPHISM-SPECIFIC POLYMERASE CHAIN REACTION AS RELIABLE AS MULTILOCUS ENZYME ELECTROPHORESIS FOR IDENTIFYING LEISHMANIA SPP. FROM ARGENTINA?

Recently, two techniques, polymerase chain reaction (PCR) amplification and sequencing of cytochrome b gene (cyt b gene sequencing) and polymorphism-specific PCR (PS-PCR) were recommended for Leishmania species identification. Before this study, however, the accuracy of these methods had not been tested against the multilocus enzyme electrophoresis, the current gold standard technique on this task. Therefore, a trial was done for the first time to compare the results obtained by these techniques, using 17 Argentinean Leishmania stocks in independent assays. For all the stocks examined, the same results at species level were obtained by the three techniques. Among them, 14 were assigned to L. (Viannia) braziliensis, and three to L. (V.) guyanensis. The two techniques, cyt b gene sequencing and PS-PCR, were able to distinguish between all the proven species responsible for leishmaniases in Argentina. Thus, both techniques were validated and could be used independently for the species designation of Leishmania parasites in the country.

Received August 10, 2005. Accepted for publication April 20, 2006.

Acknowledgments: We are grateful to Manuel Calvopiña, Hideo Kumazawa, Patricio Diosque, and Daniel Ribble for their contributions and valuable comments. We also thank Kyoko Imamura and the laboratory staffs of the Instituto de Patología Experimental and Instituto de Enfermedades Tropicales for their contributions.

Financial support: This study was supported by the Ministry of Education, Science, Culture and Sports of Japan (Grant Nos. 14256002, 15590371, and 18256004).

^* Address correspondence to Jorge D. Marco, Department of Parasitology, Kochi Medical School, Kochi University, Kohasu, Oko, Nankoku, Kochi 783-8505, Japan. E-mail: marcojd{at}med.kochi-ms.ac.jp

Authors’ addresses: Jorge D. Marco, Paola A. Barroso, Masataka Korenaga, and Yoshihisa Hashiguchi, Department of Parasitology, Kochi Medical School, Kochi University, Kohasu, Oko, Nankoku, Kochi 783-8505, Japan, Tel-Fax: +81 88 880 2415, E-mails: marcojd{at}med.kochi-ms.ac.jp, paola{at}med.kochi-u.ac.jp, korenaga{at}med.kochi-u.ac.jp, and hasiguti{at}med.kochi-u.ac.jp. Hiroshi Uezato and Shigeo Nonaka, Department of Dermatology, Faculty of Medicine, University of Ryukyus, 207 Uehara, Nishihara, Okinawa 903-0125, Japan, Tel: +81 98 895 1152, Fax: +81 98 895 1417, E-mails: huezato{at}med.u-ryukyu.ac.jp, and shigeo{at}med.u-ryukyu.ac.jp. Tat-suyuki Mimori, Department of Tumor Genetics & Biology, Graduate School of Medical Science, Kumamoto University, 1-1-1 Honjo, Ku-mamoto 860-8556, Japan, Telephone: +81 96 373 5117, Fax: +81 96 373 5120, E-mail: tmimori{at}kaiju.medic.kumamoto-u.ac.jp. Miguel A. Basombrío, Instituto de Patología Experimental, Facultad de Ciencias de la Salud, Universidad Nacional de Salta, Buenos Aires 177, (4400) Salta, Argentina, Tel-Fax: +54 387 4255333, E-mail: basombri{at}unsa.edu.ar. Néstor J. Taranto, Instituto de Enfermedades Tropicales, Sede Regional Orán, Universidad Nacional de Salta, Alvarado 751, San Ramón de la Nueva Orán, (4530) Salta, Argentina, Tel-Fax: +54 3878 421924, E-mail: inst_trop{at}arnet.com.ar.

Reprint requests: Jorge D. Marco, Department of Parasitology, Kochi Medical School, Kochi University, Kohasu, Oko, Nankoku, Kochi 783-8505, Japan. Tel-Fax: +81 88 880 2415. E-mail: marcojd{at}med.kochi-ms.ac.jp.

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