title: Molecular Characterization of Leishmania Parasites in Giemsa-Stained Slides from Cases of Human Cutaneous and Visceral Leishmaniasis, Eastern Algeria.
abstract: In Algeria, visceral leishmaniasis (VL) is due to Leishmania (L.) infantum, while three cutaneous forms (CL) are caused by Leishmania major, Leishmania tropica and Leishmania infantum. In this study, the use of Giemsa-stained slides was evaluated with two PCR techniques, in Eastern Algeria.
  
  
  
  A total of 136 samples corresponding to 100 CL smears (skin scrapings) and 36 VL slides (bone marrow aspirates) collected from 2008 to 2014 were tested. Upon DNA extraction, two PCRs were used to amplify the ribosomal Internal Transcribed Spacer 1 (ITS1) and mini-exon genes. Amplified products were digested (PCR-RFLP) and profiles analyzed for Leishmania species identification. A statistical analysis was also performed.
  
  
  
  ITS1-PCR was found significantly more sensitive than mini-exon-PCR (77.95% positives vs. 67.65%; p = 0.001). Comparison of PCR positivity showed statistically significant differences between old and recently prepared slides suggesting a better use of recent slides in PCR analyses. For species identification, PCR-restriction fragment length polymorphism (RFLP) results of ITS1 and mini-exon were concordant. L. infantum was identified from VL cases and L. infantum, L. major, and L. tropica from CL ones. According to geographical origin, L. infantum was found in North-Eastern provinces, while L. major was distributed from the North to the Center-East of Algeria. Interestingly, two L. tropica samples were identified in Annaba, located far North-East Algeria.
  
  
  
  Distribution of leishmaniasis in Eastern parts of Algeria, besides finding of L. tropica in the far North, is in this study described for the first time using molecular tools, thus confirming the usefulness of slides for PCR identification of Leishmania parasites in retrospective epidemiological investigations.
authors.affiliation: 1 Laboratoire d'Epidémiologie Moléculaire et Pathologie Expérimentale Appliquée aux Maladies Infectieuses (LR11IPT04), Institut Pasteur de Tunis, Université Tunis El Manar , Tunis, Tunisia .|2 Laboratoire de Parasitologie , CHU Annaba, Algeria .|3 Département de Biochimie, Faculté des Sciences, Université Badji Mokhtar , Annaba, Algeria .
authors.forename: Nadia
authors.initials: N
authors.lastname: Beldi
authors.affiliation: 2 Laboratoire de Parasitologie , CHU Annaba, Algeria .|4 Faculté de Médecine de Annaba, Annaba, Algeria .
authors.forename: Roukaya
authors.initials: R
authors.lastname: Mansouri
authors.affiliation: 5 Laboratoire d'Epidémiologie Médicale, Institut Pasteur de Tunis , Université Tunis El Manar, Tunis, Tunisia .
authors.forename: Jihene
authors.initials: J
authors.lastname: Bettaieb
authors.affiliation: 1 Laboratoire d'Epidémiologie Moléculaire et Pathologie Expérimentale Appliquée aux Maladies Infectieuses (LR11IPT04), Institut Pasteur de Tunis, Université Tunis El Manar , Tunis, Tunisia .
authors.forename: Alia
authors.initials: A
authors.lastname: Yaacoub
authors.affiliation: 1 Laboratoire d'Epidémiologie Moléculaire et Pathologie Expérimentale Appliquée aux Maladies Infectieuses (LR11IPT04), Institut Pasteur de Tunis, Université Tunis El Manar , Tunis, Tunisia .
authors.forename: Hejer
authors.initials: H
authors.lastname: Souguir Omrani
authors.affiliation: 1 Laboratoire d'Epidémiologie Moléculaire et Pathologie Expérimentale Appliquée aux Maladies Infectieuses (LR11IPT04), Institut Pasteur de Tunis, Université Tunis El Manar , Tunis, Tunisia .
authors.forename: Yusr
authors.initials: Y
authors.lastname: Saadi Ben Aoun
authors.affiliation: 2 Laboratoire de Parasitologie , CHU Annaba, Algeria .|4 Faculté de Médecine de Annaba, Annaba, Algeria .
authors.forename: Farida
authors.initials: F
authors.lastname: Saadni
authors.affiliation: 1 Laboratoire d'Epidémiologie Moléculaire et Pathologie Expérimentale Appliquée aux Maladies Infectieuses (LR11IPT04), Institut Pasteur de Tunis, Université Tunis El Manar , Tunis, Tunisia .
authors.forename: Ikram
authors.initials: I
authors.lastname: Guizani
authors.affiliation: 1 Laboratoire d'Epidémiologie Moléculaire et Pathologie Expérimentale Appliquée aux Maladies Infectieuses (LR11IPT04), Institut Pasteur de Tunis, Université Tunis El Manar , Tunis, Tunisia .
authors.forename: Souheila
authors.initials: S
authors.lastname: Guerbouj
mesh_terms: D000462:Algeria / Q000453:epidemiology; D000818:Animals; D001399:Azure Stains; D001853:Bone Marrow / Q000469:parasitology; D021903:DNA, Ribosomal Spacer / Q000235:genetics; D006801:Humans; D007891:Leishmania / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D016773:Leishmaniasis, Cutaneous / Q000453:epidemiology / Q000469:parasitology*; D007898:Leishmaniasis, Visceral / Q000453:epidemiology / Q000469:parasitology*; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length; D012867:Skin / Q000469:parasitology; D015047:Zoonoses
chemical_list: D001399:Azure Stains; D021903:DNA, Ribosomal Spacer
keywords: Giemsa-stained slides; ITS1; PCR-RFLP; diagnosis; leishmaniasis; mini-exon