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MULTILOCUS ENZYME ELECTROPHORESIS AND CYTOCHROME B GENE SEQUENCING–BASED IDENTIFICATION OF LEISHMANIA ISOLATES FROM DIFFERENT FOCI OF CUTANEOUS LEISHMANIASIS IN PAKISTAN

Seventeen Leishmania stocks isolated from cutaneous lesions of Pakistani patients were studied by multilocus enzyme electrophoresis and by polymerase chain reaction amplification and sequencing of the cytochrome b (Cyt b) gene. Eleven stocks that expressed nine zymodemes were assigned to L. (Leishmania) major. All of them were isolated from patients in the lowlands of Larkana district and Sibi city in Sindh and Balochistan provinces, respectively. The remaining six, distributed in two zymodemes (five and one), isolated from the highland of Quetta city, Balochistan, were identified as L. (L.) tropica. The same result at species level was obtained by the Cyt b sequencing for all the stocks examined. No clear-cut association between the clinical features (wet or dry type lesions) and the Leishmania species involved was found. Leishmania (L.) major was highly polymorphic compared with L. (L.) tropica. This difference may be explained by the fact that humans may act as a sole reservoir of L. (L.) tropica in anthroponotic cycles; however, many wild mammals can be reservoirs of L. (L.) major in zoonotic cycles.

Received October 20, 2005. Accepted for publication March 9, 2006.

Acknowledgments: The authors thank all the people and health care providers for cooperation in the study; Leandro A. Llanza for the design of the software used to calculate the Jaccard distances and the distances matrix; Patricio Diosque, Manuel Calvopiña, and Daniel Ribble for contributions and valuable comments; and Debbie Nolder for providing the Leishmania reference strains. We also thank M. Iqbal Tareen, A. Malik Tareen, Irfanullah K. Tareen, and Hashim for providing facilities and support for the examination of patients at Quetta city and Mari Abad, Quetta, Balochistan, Pakistan.

Financial support: This study was supported by Ministry of Education, Science, Culture and Sports of Japan Grants 14256002 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, 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. Abdul M. Bhutto, Doctors’ colony, Bunglow no. 14, VIP Road, Larkana, Sindh, Pakistan, E-mail: bhutto_manan{at}hotmail.com. Farooq R. Soomro and Javed H. Baloch, Incharge Leprosy Unit, CMC Hospital, Larkana, Sindh, Pakistan, E-mails: farooq_soomro5{at}hotmail.com and javedhussainb{at}yahoo.com. Hirotomo Kato, Department of Veterinary Hygiene, Faculty of Agriculture, Yamaguchi University, Yamaguchi 753-8515, Japan, E-mail: katoh{at}yamaguchi-u.ac.jp. Hiroshi Uezato and Shigeo Nonaka, Department of Dermatology, Faculty of Medicine, University of the Ryukyus, 207 Uehara, Nishihara, Okinawa 903-0125, Japan, E-mails: huezato{at}med.u-ryukyu.ac.jp and shigeo{at}med.u-ryukyu.ac.jp. Ken Katakura, Department of Disease Control, Graduate School of Veterinary Medicine, Hokkaido University, Kita 18 Nishi 9, Kita, Sapporo 060-0818, Japan, E-mail: kenkata{at}vetmed.hokudai.ac.jp.

Reprint requests: 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.

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