title: Comparison of Leishmania typing results obtained from 16 European clinical laboratories in 2014.
abstract: Leishmaniasis is endemic in southern Europe, and in other European countries cases are diagnosed in travellers who have visited affected areas both within the continent and beyond. Prompt and accurate diagnosis poses a challenge in clinical practice in Europe. Different methods exist for identification of the infecting Leishmania species. Sixteen clinical laboratories in 10 European countries, plus Israel and Turkey, conducted a study to assess their genotyping performance. DNA from 21 promastigote cultures of 13 species was analysed blindly by the routinely used typing method. Five different molecular targets were used, which were analysed with PCR-based methods. Different levels of identification were achieved, and either the Leishmania subgenus, species complex, or actual species were reported. The overall error rate of strains placed in the wrong complex or species was 8.5%. Various reasons for incorrect typing were identified. The study shows there is considerable room for improvement and standardisation of Leishmania typing. The use of well validated standard operating procedures is recommended, covering testing, interpretation, and reporting guidelines. Application of the internal transcribed spacer 1 of the rDNA array should be restricted to Old World samples, while the heat-shock protein 70 gene and the mini-exon can be applied globally.
authors.affiliation: Biomedical Sciences, Institute of Tropical Medicine, Antwerp, Belgium.
authors.forename: Gert
authors.initials: G
authors.lastname: Van der Auwera
authors.affiliation: Academic Medical Center, Amsterdam, The Netherlands.
authors.forename: Aldert
authors.initials: A
authors.lastname: Bart
authors.affiliation: Instituto de Salud Carlos III, Madrid, Spain.
authors.forename: Carmen
authors.initials: C
authors.lastname: Chicharro
authors.affiliation: Global Health and Tropical Medicine, GHTM, Instituto de Higiene e Medicina Tropical, UNL, Lisbon, Portugal.
authors.forename: Sofia
authors.initials: S
authors.lastname: Cortes
authors.affiliation: The Public Health Agency of Sweden, Stockholm, Sweden.
authors.forename: Leigh
authors.initials: L
authors.lastname: Davidsson
authors.affiliation: Istituto Superiore di Sanità, Rome, Italy.
authors.forename: Trentina
authors.initials: T
authors.lastname: Di Muccio
authors.affiliation: Biomedical Sciences, Institute of Tropical Medicine, Antwerp, Belgium.|Biomedical Sciences, Antwerp University, Antwerp, Belgium.
authors.forename: Jean-Claude
authors.initials: JC
authors.lastname: Dujardin
authors.affiliation: Swiss Tropical and Public Health Institute, Basel, Switzerland.|University of Basel, Basel, Switzerland.
authors.forename: Ingrid
authors.initials: I
authors.lastname: Felger
authors.affiliation: National Institute for Infectious Diseases (INMI) Lazzaro Spallanzani, Rome, Italy.
authors.forename: Maria Grazia
authors.initials: MG
authors.lastname: Paglia
authors.affiliation: Institute of Parasitology, University of Zürich, Zürich, Switzerland.
authors.forename: Felix
authors.initials: F
authors.lastname: Grimm
authors.affiliation: Institute of Tropical Medicine and International Health, Charité-Universitätsmedizin Berlin, Berlin, Germany.
authors.forename: Gundel
authors.initials: G
authors.lastname: Harms
authors.affiliation: Hebrew University, Hadassah Medical Centre, Jerusalem, Israel.
authors.forename: Charles L
authors.initials: CL
authors.lastname: Jaffe
authors.affiliation: United Kingdom National External Quality Assessment Service, London, United Kingdom.
authors.forename: Monika
authors.initials: M
authors.lastname: Manser
authors.affiliation: University of Montpellier, Montpellier, France.
authors.forename: Christophe
authors.initials: C
authors.lastname: Ravel
authors.affiliation: Centre Hospitalier Universitaire de Rennes, Rennes, France.
authors.forename: Florence
authors.initials: F
authors.lastname: Robert-Gangneux
authors.affiliation: National Institute for Public Health and the Environment, RIVM, Bilthoven, The Netherlands.
authors.forename: Jeroen
authors.initials: J
authors.lastname: Roelfsema
authors.affiliation: Ege University, Faculty of Medicine, Department of Parasitology, Izmir, Turkey.
authors.forename: Seray
authors.initials: S
authors.lastname: Töz
authors.affiliation: St. Elisabeth Hospital, Tilburg, The Netherlands.
authors.forename: Jaco J
authors.initials: JJ
authors.lastname: Verweij
authors.affiliation: Hospital for Tropical Diseases, London, United Kingdom.|London School of Hygiene and Tropical Medicine, London, United Kingdom.
authors.forename: Peter L
authors.initials: PL
authors.lastname: Chiodini
mesh_terms: D018105:DNA, Kinetoplast; D016054:DNA, Protozoan / Q000235:genetics; D004275:DNA, Ribosomal; D005060:Europe; D005838:Genotype; D018840:HSP70 Heat-Shock Proteins / Q000235:genetics*; D006801:Humans; D007557:Israel; D007753:Laboratories; D007891:Leishmania / Q000235:genetics* / Q000302:isolation & purification; D007896:Leishmaniasis / Q000175:diagnosis*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012150:Polymorphism, Restriction Fragment Length; D012680:Sensitivity and Specificity; D014421:Turkey
chemical_list: D018105:DNA, Kinetoplast; D016054:DNA, Protozoan; D004275:DNA, Ribosomal; D018840:HSP70 Heat-Shock Proteins
keywords: Typing; hsp70; kinetoplast DNA; leishmaniasis; mini-exon; rDNA ITS1