title: Multilocus microsatellite typing of Leishmania infantum isolates in monitored Leishmania/HIV coinfected patients.
abstract: BACKGROUND
  Leishmania infantum is the main etiological agent of both visceral and cutaneous clinical forms of leishmaniasis in the Mediterranean area. Leishmania/HIV coinfection in this area is characterized by a chronic course and frequent recurrences of clinical episodes. The present study using Multilocus Microsatellite Typing (MLMT) analysis, a highly discriminative tool, aimed to genetically characterize L. infantum isolates taken from monitored Leishmania/HIV coinfected patients presenting successive clinical episodes.
  
  
  METHODS
  In this study, by the analysis of 20 microsatellite loci, we studied the MLMT profiles of 25 L. infantum isolates from 8 Leishmania/HIV coinfected patients who had experienced several clinical episodes. Two to seven isolates per patient were taken before and after treatment, during clinical and non-clinical episodes, with time intervals of 6 days to 29 months. Genetic diversity, clustering and phenetic analyses were performed.
  
  
  RESULTS
  MLMT enabled us to study the genetic characteristics of the 25 L. infantum isolates, differentiating 18 genotypes, corresponding to a genotypic diversity of 0.72. Fifteen genotypes were unique in the total sample set and only 3 were repeated, 2 of which were detected in different patients. Both clustering and phylogenetic analyses provided insights into the genetic links between the isolates; in five patients isolates showed clear genetic links: either the genotype was exactly the same or only slightly different. In contrast, the isolates of the other three patients were dispersed in different clusters and some could be the result of mixing between populations.
  
  
  CONCLUSIONS
  Our data indicated a great MLMT variability between isolates from coinfected patients and no predominant genotype was observed. Despite this, almost all clinical episodes could be interpreted as a relapse rather than a reinfection. The results showed that diverse factors like an intrapatient evolution over time or culture bias could influence the parasite population detected in the patient, making it difficult to differentiate between relapse and reinfection.
authors.affiliation: Laboratory of Parasitology, Faculty of Pharmacy, Universitat de Barcelona, Avda Joan XXIII s/n, 08028, Barcelona, Spain. mtomasperez@outlook.com.
authors.forename: Míriam
authors.initials: M
authors.lastname: Tomás-Pérez
authors.affiliation: MIVEGEC, UMR IRD 224-CNRS 5290-Université de Montpellier, Montpellier, France. mallorie.hide@ird.fr.
authors.forename: Mallorie
authors.initials: M
authors.lastname: Hide
authors.affiliation: Laboratory of Parasitology, Faculty of Pharmacy, Universitat de Barcelona, Avda Joan XXIII s/n, 08028, Barcelona, Spain. mcriera@ub.edu.
authors.forename: Cristina
authors.initials: C
authors.lastname: Riera
authors.affiliation: Laboratory of Parasitology, Faculty of Pharmacy, Universitat de Barcelona, Avda Joan XXIII s/n, 08028, Barcelona, Spain. lilimon1@hotmail.com.
authors.forename: Liliana
authors.initials: L
authors.lastname: Montoya
authors.affiliation: MIVEGEC, UMR IRD 224-CNRS 5290-Université de Montpellier, Montpellier, France. anne-laure.banuls@ird.fr.
authors.forename: Anne-Laure
authors.initials: AL
authors.lastname: Bañuls
authors.affiliation: Infectious Diseases Department and Microbiology Department, Hospital Universitari Vall d'Hebron, Universitat Autònoma de Barcelona, Barcelona, Spain. eribera@vhebron.net.
authors.forename: Esteve
authors.initials: E
authors.lastname: Ribera
authors.affiliation: Laboratory of Parasitology, Faculty of Pharmacy, Universitat de Barcelona, Avda Joan XXIII s/n, 08028, Barcelona, Spain. mportus@ub.edu.
authors.forename: Montserrat
authors.initials: M
authors.lastname: Portús
authors.affiliation: Laboratory of Parasitology, Faculty of Pharmacy, Universitat de Barcelona, Avda Joan XXIII s/n, 08028, Barcelona, Spain. rfisa@ub.edu.
authors.forename: Roser
authors.initials: R
authors.lastname: Fisa
mesh_terms: D000328:Adult; D060085:Coinfection / Q000469:parasitology* / Q000821:virology; D005260:Female; D014644:Genetic Variation; D015658:HIV Infections / Q000821:virology*; D006801:Humans; D018314:Leishmania infantum / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification*; D007898:Leishmaniasis, Visceral / Q000469:parasitology*; D008297:Male; D018895:Microsatellite Repeats; D058885:Multilocus Sequence Typing / Q000379:methods*; D010802:Phylogeny