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Journal of Clinical Microbiology, July 2003, p. 3147-3153, Vol. 41, No. 7
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.7.3147-3153.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Identification and Differentiation of Leishmania Species in Clinical Samples by PCR Amplification of the Miniexon Sequence and Subsequent Restriction Fragment Length Polymorphism Analysis

Jutta Marfurt,1 Abed Nasereddin,2 Igor Niederwieser,1 Charles L. Jaffe,2 Hans-Peter Beck,1 and Ingrid Felger1*

Department of Medical Parasitology and Infection Biology, Swiss Tropical Institute, Basel, Switzerland,1 Department of Parasitology, Hadassah Medical School, Hebrew University, Jerusalem, Israel2

Received 19 November 2002/ Returned for modification 12 January 2003/ Accepted 28 April 2003

We recently developed a new PCR-restriction fragment length polymorphism (RFLP)-based assay using the miniexon sequence from the genus Leishmania. Here we report the application of this new genotyping method to naturally infected clinical samples for the differentiation of New and Old World Leishmania species. Of the newly developed assay and four currently applied diagnostic tests (i.e., in vitro cultivation, serology, and two other molecular assays using either the small subunit-internal transcribed spacer sequence or a repetitive genomic sequence), the miniexon assay showed the highest sensitivity, 89.7%, compared to 70.6, 57.1, 51.7, and 79.3%, respectively. Species differentiation was robust and reliable compared with that by two other Leishmania genotyping techniques. The assay provides a valuable tool for the identification of Leishmania directly from clinical samples and enables determination of the infecting species by a facile technique with high discrimination power. Since Leishmania causes a broad spectrum of diseases distinguished by different parasite and host factors, detection and characterization of the infecting species is crucial for the confirmation of a diagnosis as well as the establishment of the clinical prognosis and the initiation of an adequate therapeutic approach. The miniexon PCR-RFLP assay will facilitate such determination and might improve diagnosis and treatment of leishmaniasis.

* Corresponding author. Mailing address: Swiss Tropical Institute, Socinstrasse 57, Postfach, CH-4002 Basel, Switzerland. Phone: 41 61 284 81 17. Fax: 41 61 271 86 54. E-mail: ingrid.felger{at}unibas.ch.

Journal of Clinical Microbiology, July 2003, p. 3147-3153, Vol. 41, No. 7
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.7.3147-3153.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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