title: [Contribution of Leishmania identification using polymerase chain reaction--restriction fragment length polymerase for epidemiological studies of cutaneous leishmaniasis in Tunisia].
abstract: OBJECTIVE
  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.
  
  
  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.
  
  
  CONCLUSIONS
  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.
authors: Laboratoire de recherche LR 05-SP 03 « parasitoses émergentes », institut Pasteur de Tunis, 13, place Pasteur, BP74, 1002 Tunis, Tunisie.
authors: N
authors: N
authors: Bousslimi
authors: Laboratoire de recherche LR 05-SP 03 « parasitoses émergentes », institut Pasteur de Tunis, 13, place Pasteur, BP74, 1002 Tunis, Tunisie; Laboratoire de parasitologie-mycologie, institut Pasteur de Tunis, 13, place Pasteur, BP74, 1002 Tunis, Tunisie. Electronic address: benabda.imen@hotmail.fr.
authors: I
authors: I
authors: Ben Abda
authors: Laboratoire de recherche LR 05-SP 03 « parasitoses émergentes », institut Pasteur de Tunis, 13, place Pasteur, BP74, 1002 Tunis, Tunisie; Service de dermatologie, hôpital Habib Thameur, 8, rue Ali Ben Ayed Montfleury, 1008 Tunis, Tunisie.
authors: R
authors: R
authors: Ben Mously
authors: Laboratoire de recherche LR 05-SP 03 « parasitoses émergentes », institut Pasteur de Tunis, 13, place Pasteur, BP74, 1002 Tunis, Tunisie; Laboratoire de parasitologie-mycologie, institut Pasteur de Tunis, 13, place Pasteur, BP74, 1002 Tunis, Tunisie.
authors: E
authors: E
authors: Siala
authors: Laboratoire d'épidémiologie parasitaire, institut Pasteur d'Algérie, 2, route du Petit Staouèli Dély Ibrahim, 16047 Alger, Algérie.
authors: Z
authors: Z
authors: Harrat
authors: Laboratoire de parasitologie-mycologie, institut Pasteur de Tunis, 13, place Pasteur, BP74, 1002 Tunis, Tunisie.
authors: N
authors: N
authors: Zallagua
authors: Laboratoire de recherche LR 05-SP 03 « parasitoses émergentes », institut Pasteur de Tunis, 13, place Pasteur, BP74, 1002 Tunis, Tunisie; Laboratoire de parasitologie-mycologie, institut Pasteur de Tunis, 13, place Pasteur, BP74, 1002 Tunis, Tunisie.
authors: A
authors: A
authors: Bouratbine
authors: Laboratoire de recherche LR 05-SP 03 « parasitoses émergentes », institut Pasteur de Tunis, 13, place Pasteur, BP74, 1002 Tunis, Tunisie; Laboratoire de parasitologie-mycologie, institut Pasteur de Tunis, 13, place Pasteur, BP74, 1002 Tunis, Tunisie.
authors: K
authors: K
authors: Aoun
mesh_terms: D016054:DNA, Protozoan / Q000032:analysis / Q000235:genetics; D021903:DNA, Ribosomal Spacer / Q000032:analysis / Q000235:genetics; D006801:Humans; D007527:Isoenzymes / Q000032:analysis; D007891:Leishmania / Q000201:enzymology / Q000235:genetics / Q000302:isolation & purification*; D018314:Leishmania infantum / Q000201:enzymology / Q000235:genetics / Q000302:isolation & purification; D018320:Leishmania major / Q000201:enzymology / Q000235:genetics / Q000302:isolation & purification; D007895:Leishmania tropica / Q000201:enzymology / Q000235:genetics / Q000302:isolation & purification; D016773:Leishmaniasis, Cutaneous / Q000453:epidemiology / Q000469:parasitology*; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length; D015800:Protozoan Proteins / Q000032:analysis; D015203:Reproducibility of Results; D012867:Skin / Q000469:parasitology; D013045:Species Specificity; D014416:Tunisia / Q000453:epidemiology
chemical_list: D016054:DNA, Protozoan; D021903:DNA, Ribosomal Spacer; D007527:Isoenzymes; D015800:Protozoan Proteins
keywords: Cutaneous leishmaniasis; Isoenzyme electrophoresis; Leishmania; Leishmaniose cutanée; PCR-RFLP; Tunisia; Tunisie; Électrophorèse des isoenzymes