TY - JOUR
T1 - Species-specific markers for the differential diagnosis of Trypanosoma cruzi and Trypanosoma rangeli and polymorphisms detection in Trypanosoma rangeli
AU - Ferreira, Keila Adriana Magalhães
AU - Fajardo, Emanuella Francisco
AU - Baptista, Rodrigo P.
AU - Macedo, Andrea Mara
AU - Lages-Silva, Eliane
AU - Ramírez, Luis Eduardo
AU - Pedrosa, André Luiz
PY - 2014/6
Y1 - 2014/6
N2 - Trypanosoma cruzi and Trypanosoma rangeli are kinetoplastid parasites which are able to infect humans in Central and South America. Misdiagnosis between these trypanosomes can be avoided by targeting barcoding sequences or genes of each organism. This work aims to analyze the feasibility of using species-specific markers for identification of intraspecific polymorphisms and as target for diagnostic methods by PCR. Accordingly, primers which are able to specifically detect T. cruzi or T. rangeli genomic DNA were characterized. The use of intergenic regions, generally divergent in the trypanosomatids, and the serine carboxypeptidase gene were successful. Using T. rangeli genomic sequences for the identification of group-specific polymorphisms and a polymorphic AT(n) dinucleotide repeat permitted the classification of the strains into two groups, which are entirely coincident with T. rangeli main lineages, KP1 (+) and KP1 (-), previously determined by kinetoplast DNA (kDNA) characterization. The sequences analyzed totalize 622 bp (382 bp represent a hypothetical protein sequence, and 240 bp represent an anonymous sequence), and of these, 581 (93.3 %) are conserved sites and 41 bp (6.7 %) are polymorphic, with 9 transitions (21.9 %), 2 transversions (4.9 %), and 30 (73.2 %) insertion/deletion events. Taken together, the species-specific markers analyzed may be useful for the development of new strategies for the accurate diagnosis of infections. Furthermore, the identification of T. rangeli polymorphisms has a direct impact in the understanding of the population structure of this parasite.
AB - Trypanosoma cruzi and Trypanosoma rangeli are kinetoplastid parasites which are able to infect humans in Central and South America. Misdiagnosis between these trypanosomes can be avoided by targeting barcoding sequences or genes of each organism. This work aims to analyze the feasibility of using species-specific markers for identification of intraspecific polymorphisms and as target for diagnostic methods by PCR. Accordingly, primers which are able to specifically detect T. cruzi or T. rangeli genomic DNA were characterized. The use of intergenic regions, generally divergent in the trypanosomatids, and the serine carboxypeptidase gene were successful. Using T. rangeli genomic sequences for the identification of group-specific polymorphisms and a polymorphic AT(n) dinucleotide repeat permitted the classification of the strains into two groups, which are entirely coincident with T. rangeli main lineages, KP1 (+) and KP1 (-), previously determined by kinetoplast DNA (kDNA) characterization. The sequences analyzed totalize 622 bp (382 bp represent a hypothetical protein sequence, and 240 bp represent an anonymous sequence), and of these, 581 (93.3 %) are conserved sites and 41 bp (6.7 %) are polymorphic, with 9 transitions (21.9 %), 2 transversions (4.9 %), and 30 (73.2 %) insertion/deletion events. Taken together, the species-specific markers analyzed may be useful for the development of new strategies for the accurate diagnosis of infections. Furthermore, the identification of T. rangeli polymorphisms has a direct impact in the understanding of the population structure of this parasite.
KW - Differential diagnosis
KW - Genetic polymorphisms
KW - Microsatellites
KW - PCR
KW - Trypanosoma cruzi
KW - Trypanosoma rangeli
UR - http://www.scopus.com/inward/record.url?scp=84903816597&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84903816597&partnerID=8YFLogxK
U2 - 10.1007/s00436-014-3872-2
DO - 10.1007/s00436-014-3872-2
M3 - Article
C2 - 24728520
AN - SCOPUS:84903816597
SN - 0932-0113
VL - 113
SP - 2199
EP - 2207
JO - Parasitology Research
JF - Parasitology Research
IS - 6
ER -