Heat-shock protein 70 PCR-RFLP: a universal simple tool for Leishmania species discrimination in the New and Old World

AM Montalvo, J Fraga, L Monzote, I Montano, S De Doncker, JC Dujardin, G Van der Auwera

    Research output: Contribution to journalA1: Web of Science-article

    Abstract

    SUMMARYIntroduction. Species typing in leishmaniasis gains importance in diagnostics, epidemiology, and clinical studies. A restriction fragment length polymorphism (RFLP) assay of PCR amplicons from a partial heat-shock protein 70 gene (hsp70) had been described for the New World, allowing identification of some species. Methods. Based on an initial in silico analysis of 51 hsp70 sequences, most of which we recently determined in the frame of a phylogenetic study, species-specific restriction sites were identified. These were tested by PCR-RFLP on 139 strains from 14 species, thereby documenting both inter- and intra-species variability. Results. Our assay could identify Leishmania infantum, L. donovani, L. tropica, L. aethiopica, L. major, L. lainsoni, L. naiffi, L. braziliensis, L. peruviana, L. guyanensis, and L. panamensis by applying 2 subsequent digests. L. mexicana, L. amazonensis, and L. garnhami did not generate species-specific restriction fragment patterns. Conclusion. Currently no assay is available for global Leishmania species discrimination. We present a universal PCR-RFLP method allowing identification of most medically relevant Old and New World Leishmania species on the basis of a single PCR, obviating the need to perform separate PCRs. The technique is simple to perform and can be implemented in all settings where PCR is available
    Original languageEnglish
    JournalParasitology
    Volume137
    Issue number8
    Pages (from-to)1159-1168
    Number of pages10
    ISSN0031-1820
    DOIs
    Publication statusPublished - 2010

    Keywords

    • B780-tropical-medicine
    • Protozoal diseases
    • Leishmaniasis
    • Leishmania infantum
    • Leishmania donovani
    • Leishmania tropica
    • Leishmania aethiopica
    • Leishmania major
    • Leishmania lainsoni
    • Leishmania naiffi
    • Leishmania braziliensis
    • Leishmania peruviana
    • Leishmania guyanensis
    • Leishmania panamensis
    • Differential diagnosis
    • Identification
    • Species
    • Strains
    • Polymerase chain reaction
    • PCR-RFLP
    • Efficacy
    • Laboratory techniques and procedures

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