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. 2017 May 12;10(1):236.
doi: 10.1186/s13071-017-2175-8.

Evaluation of a Multilocus Sequence Typing (MLST) scheme for Leishmania (Viannia) braziliensis and Leishmania (Viannia) panamensis in Colombia

Giovanny Herrera  1 , Carolina Hernández  1 , Martha S Ayala  2 , Carolina Flórez  2 , Aníbal A Teherán  3 , Juan David Ramírez  4
Affiliations

Evaluation of a Multilocus Sequence Typing (MLST) scheme for Leishmania (Viannia) braziliensis and Leishmania (Viannia) panamensis in Colombia

Giovanny Herrera et al. Parasit Vectors. .

Abstract

Background: Leishmaniases are parasitic vector-borne diseases affecting more than 12 million people in 98 countries. In Colombia, leishmaniasis is widespread and the most common clinical manifestation is cutaneous, mainly caused by L. panamensis and L. braziliensis. Currently, the genetic diversity of these species in Colombia is unknown. To address this, we applied molecular techniques for their characterization, using multilocus sequence typing (MLST) to explore the genetic variability and phylodynamics of the disease.

Methods: Seven previously described genetic markers were selected highlighting the implementation of a mitochondrial marker. Markers were applied to 163 samples from isolates obtained between 1980 and 2001.

Results: The identification of the samples showed an excellent correlation with typing tests previously applied (MLEE, monoclonal antibodies). Isolates of L. braziliensis showed greater genetic diversity than L. panamensis, and a greater number of diploid sequence types (DSTs). In addition, the geographical distribution of DSTs for each species were obtained through georeferencing maps.

Conclusions: To our knowldge, this study represents the first description of the genetic variability of L. panamensis in Colombia and South America, and is the first to propose a scheme of MLST for epidemiological surveillance of leishmaniasis in the country.

Keywords: Cutaneous leishmaniasis; DST; Genetic diversity; Leishmania; MLST.

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Figures

Fig. 1
Fig. 1
Comparison of genetic diversity indexes (Hd, π and θ) by geographical region for L. panamensis (a) and L. braziliensis (b)
Fig. 2
Fig. 2
Geographical distribution of diploid sequence types (DSTs) retrieved based on the seven gene markers MLST scheme for L. braziliensis (a) and L. panamensis (b)
Fig. 3
Fig. 3
Temporal variation of diploid sequence types (DSTs) retrieved based on the seven gene markers MLST scheme for L. panamensis (a) and L. braziliensis (b)
Fig. 4
Fig. 4
Phylogenetic reconstruction based on the seven gene markers MLST scheme. a Maximum likelihood phylogenetic reconstruction cladogram of L. panamensis (red) and L. braziliensis (blue) of concatenated sequences of the seven markers employed sowing representatives of the DSTs detected. b Clonal complex analysis performed on eBURST showing the number of clonal complexes for L. braziliensis (blue) and L. panamensis (red) using the seven gene markers of the MLST scheme
Fig. 5
Fig. 5
Allelic plot based on the seven gene markers MLST scheme. Allelic plot showing the inheritance of alleles for L. braziliensis (blue) and L. panamensis (red) based on phylogenetic reconstruction and considering a true allele with a bootstrap cut-off equal or over 80%. The different shades of blue/red indicate variants of alleles supported by bootstrap (over or equal 80%) within each species studied
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