The compositional landscape of minicircle sequences isolated from active lesions and scars of American cutaneous leishmaniasis.
BACKGROUND
American cutaneous leishmaniasis (ACL) is characterized by cutaneous lesions that heal spontaneously or after specific treatment. This paper reports on the analysis of kDNA minicircle sequences from clinical samples (typical lesions and scars) that were PCR-amplified with specific primers for Leishmania species of the subgenus Viannia.


METHODS
From 56 clinical isolates we obtained a single amplified fragment (ca. 790 bp), which after cloning and sequencing resulted in 290 minicircle sequences from both active lesions and scars. We aimed to get a compositional profile of these sequences in clinical samples and evaluate the corresponding compositional changes. Sequences were analyzed with the compseq and wordcount (Emboss package) to get the composition of di-, tri-, tetra-, penta- and hexanucleotides. Additionally, we built a nucleotide dictionary with words of 7, 8, 9 and 10 nucleotides.


RESULTS
This compositional analysis showed that minicircles amplified from active cutaneous lesions and scars have a distinct compositional profile as viewed by nucleotide composition of words up to 10mer. With regard to the most frequent nucleotide words above length 6, there is also a distinct pattern for 7, 8, 9 and 10mer.


CONCLUSIONS
These results indicate that minicircle sequences can be monitored upon direct exposure to a selection/stressing environment (e.g. chemical action) by evaluating their nucleotide compositional profile. It might be useful as a molecular tool in research concerning the evolution of infecting Leishmania in both vector and vertebrate hosts.
Eduardo Henrique Gomes
EH
Rodrigues
Fábia Carla da Silva
FC
Soares
Roberto Pereira
RP
Werkhäuser
Maria Edileuza F
ME
de Brito
Octavio
O
Fernandes
Frederico G Coutinho
FG
Abath
Adeilton
A
Brandão
D001483:Base Sequence; D017931:DNA Primers / Q000235:genetics; D018105:DNA, Kinetoplast / Q000235:genetics*; D006801:Humans; D007891:Leishmania / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification; D016773:Leishmaniasis, Cutaneous / Q000469:parasitology*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA
D017931:DNA Primers; D018105:DNA, Kinetoplast