title: Intraspecific genetic variability in a population of Moroccan Leishmania infantum revealed by PCR-RFLP of kDNA minicircles.
abstract: In Morocco, Leishmania infantum is the main etiologic agent of human and canine visceral leishmaniasis (VL). This species has been proven to be an opportunistic agent in HIV+ patients and is also responsible of sporadic cutaneous leishmaniasis (CL).This work aims to evaluate the genetic variability of Moroccan L. infantum strains based on PCR-RFLP analysis of the kinetoplastid DNA (kDNA) minicircles. A total of 75 DNA samples extracted from positive Giemsa-stained smears (n=32) and from L. infantum cultures (n=43) was studied. The samples have been taken from VL patients infected (n=7) or not (n=56) by HIV, patients with CL (n=2) and finally from infected dogs (n=10). An hypervariable region of kDNA was amplified using the primers MC1 and MC2; the PCR products were digested separately by a panel of nine restriction enzymes. The presence or absence of restriction fragments was scored in a binary matrix and the SplitsTree4 software was used for the construction of a Neighbor-Net network. Moroccan L. infantum population showed an important level of variability with the identification of 6 genotypes. For each genotype a PCR product was sequenced, confirming the presence of all the expected restriction sites. The predominant profile was the genotype B. A new genotype, named Q was detected for the first time, whereas the four other genotypes (G, K, N and O) were reported sporadically in the Mediterranean basin. The Neighbor-Net network segregates our L. infantum population into 3 clusters: Cluster I includes genotype B, cluster II grouping the genotypes O, Q and G and finally the cluster III contains the genotype N. The kDNA-PCR-RFLP assay is suitable for use directly on biological samples; it reveals an important degree of genetic variability among L. infantum strains even those belonging to the same zymodeme what is of great epidemiological interest.
authors.affiliation: Laboratory of Parasitology and Vector-Borne-Diseases, Institut Pasteur du Maroc, Casablanca, Morocco.
authors.forename: Adil
authors.initials: A
authors.lastname: El Hamouchi
authors.affiliation: Laboratory of Parasitology and Vector-Borne-Diseases, Institut Pasteur du Maroc, Casablanca, Morocco.
authors.forename: Rajaa
authors.initials: R
authors.lastname: Ejghal
authors.affiliation: Faculty of Medicine and Pharmacy, University Sidi Mohammed Ben Abdellah, Route de Sefrou, Fes, Morocco.
authors.forename: Moustapha
authors.initials: M
authors.lastname: Hida
authors.affiliation: Laboratory of Parasitology and Vector-Borne-Diseases, Institut Pasteur du Maroc, Casablanca, Morocco. Electronic address: meryem.lemrani@pasteur.ma.
authors.forename: Meryem
authors.initials: M
authors.lastname: Lemrani
mesh_terms: D000818:Animals; D001483:Base Sequence; D017931:DNA Primers; D018105:DNA, Kinetoplast / Q000235:genetics; D004283:Dog Diseases / Q000453:epidemiology / Q000382:microbiology*; D004285:Dogs; D005838:Genotype; D015658:HIV Infections / Q000150:complications / Q000382:microbiology*; D006801:Humans; D018314:Leishmania infantum / Q000235:genetics*; D007898:Leishmaniasis, Visceral / Q000150:complications / Q000453:epidemiology / Q000382:microbiology*; D009018:Morocco; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length
chemical_list: D017931:DNA Primers; D018105:DNA, Kinetoplast
keywords: Genetic variability; Kinetoplastid DNA; Leishmania infantum; Morocco; PCR-RFLP