1. Place de l’identification des leishmanies par la polymerase chain reaction – restriction fragment length polymerase dans l’étude de l’épidémiologie des leishmanioses cutanées en Tunisie
- Author
-
Bousslimi, N., Ben Abda, I., Ben Mously, R., Siala, E., Harrat, Z., Zallagua, N., Bouratbine, A., Aoun, K., Laboratoire des parasitoses émergentes, Institut Pasteur de Tunis (LR 05-SP 03), Institut Pasteur de Tunis, Réseau International des Instituts Pasteur (RIIP)-Réseau International des Instituts Pasteur (RIIP), Laboratoire de parasitologie-mycologie, parasitoses médicales, biotechnologie et biomolécules, Service de dermatologie, hôpital Habib Thameur, 8, rue Ali Ben Ayed Montfleury, 1008 Tunis, Tunisie, Institut Pasteur d'Algérie, Réseau International des Instituts Pasteur (RIIP), and Ce travail a bénéficié du soutien du ministère tunisien de l’Enseignement Supérieur et de la Recherche Scientifique dans le cadre du financement du LR 11-IPT-06 et des actions concertées inter-pasteuriennes dans le cadre du projet ACIP A 04-2007
- Subjects
Leishmania ,Cutaneous leishmaniasis ,Tunisia ,[SDV]Life Sciences [q-bio] ,Protozoan Proteins ,Leishmaniasis, Cutaneous ,Reproducibility of Results ,Isoenzyme electrophoresis ,DNA, Protozoan ,Polymerase Chain Reaction ,Tunisie ,Isoenzymes ,PCR-RFLP ,Leishmaniose cutanée ,[SDV.MP]Life Sciences [q-bio]/Microbiology and Parasitology ,Species Specificity ,Leishmania tropica ,DNA, Ribosomal Spacer ,Electrophorèse des isoenzymes ,Humans ,Leishmania infantum ,Polymorphism, Restriction Fragment Length ,Leishmania major ,Skin - Abstract
International audience; AIM: Three forms of cutaneous leishmaniasis (CL) are endemic in Tunisia. The identification of the causative species is useful to complete epidemiological data and to manage the cases. The aim of this study is to assess PCR-RFLP technique in the identification of Leishmania species responsible of CL in Tunisia and to compare the results of this technique to those of isoenzyme analysis. PATIENTS AND METHODS: Sixty-one CL lesions were sampled. Dermal samples were tested by culture on NNN medium and analyzed by PCR-RFLP assay targeting the ITS1 region of ribosomal DNA. Species identification was performed by both iso-enzymatic typing for positive cultures and analysis of restriction profiles after enzymatic digestion by HaeIII of the obtained amplicons. RESULTS: Thirty-eight (62%) samples were positive by culture. The iso-enzymatic typing of 32 isolates identified 3 L. infantum, 23 L. major MON-25 and 6 L. tropica MON-8. Sixty samples were positive by PCR. The PCR-RFLP digestion profiles of the 56 PCR products identified 12 L. infantum, 38 L. major and 6 L. tropica. The results of both techniques were concordant in the 32 strains identified by both techniques. Species identification correlated with the geographical distribution of CL forms endemic in Tunisia. CONCLUSION: Results of PCR-RFLP revealed highly concordant with those of isoenzyme electrophoresis. Thanks to its simplicity, rapidity and ability to be performed directly on biological samples, this technique appears as an interesting alternative for the identification of Leishmania strains responsible of CL in Tunisia.
- Published
- 2012
- Full Text
- View/download PDF