First approach for a consensus multilocus sequence typing scheme for worldwide Leishmania spp. identification

LAUTHIER JUAN JOSÉ; RUYBAL PAULA; BARROSO PAOLA; HOYOS CARLOS; PORTELLI SABRINA; BRACAMONTE MARIA; LOCATELLI FABRICIO; HASHIGUCHI YOSHIHISA; MARCO JORGE DIEGO; KORENAGA MASATAKA

Osaka

Encuentro; The 56th Annual Meeting for the Japanese Society of Tropical Medicine; 2015

Japanese Society of Tropical Medicine

Leishmaniasis is one of the world?s most neglected diseases, affecting the poorest populations in most tropical and subtropical regions. The causative agent corresponds to flagellated protozoan parasites of the genus Leishmania. About 21 species are known to infect humans. Multilocus Sequence Typing (MLST) is a method based on nucleotide polymorphism analysis of internal fragments of housekeeping genes. In the present work we propose a first in-silico approach for a consensus MLST scheme for Leishmania spp., using available genomic data. Twenty-seven gene fragments, previously published by other authors, for 18 Leishmania spp. were studied. A total of 33,474 bp were analyzed and 35.21% of polymorphisms in 11,789 sites. The highest alleles number and discriminatory power (DP) by diploid sequence types (DST) analysis were 6PGD, ME, HX, ENO, Ch28 and Ch36-1190 genes (18 alleles and 0.995 DP respectively), meanwhile LACK gene shown the lowest values (13 alleles and 0.963 DP). The global DP was 0,994. A scheme optimization was carried out by testing all possible combinations among the genes (MLSTest 1.0). Monophyly of the groups and branch support were maximized. To become the new gold standard for Leishmania spp. typing, we select 15 gene candidates (ASAT, GPI, 6PGD, PPA, MKK, ICD, MPI, G6PD, PGM, HX, ACON, ALAT, ENO, PMM and Ch36-1190) that had the best conditions, monophyly and good branch support for every complex and species. To date, this is the first attempt to obtain a consensus MLST scheme from all the proposed gene targets.