Abstract
BackgroundAmerican 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.MethodsFrom 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.ResultsThis 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.ConclusionThese 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.
Highlights
American cutaneous leishmaniasis (ACL) is characterized by cutaneous lesions that heal spontaneously or after specific treatment
American cutaneous leishmaniasis (ACL) is a zoonotic disease caused by Leishmania (Viannia) spp., L. (Leishmania) amazonensis/mexicana or L. (L.) infantum, a protozoan parasite which infects the vertebrate host after being bitten by infected phlebotomus insects of the genus Lutzomya
We show here the results of such a compositional analysis of Leishmania (V.) spp. minicircle sequences from ACL patients coming from endemic regions in Pernambuco state, Brazil
Summary
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. The work of Karlin and Mrázek [14] demonstrated that the nucleotide composition of a particular species is biased toward some of the sixteen possible dinucleotides. This bias can be viewed as a peculiar genome signature and under certain assumptions the dinucleotide bias might reveal evolutionary distance [14]. Some studies have reported on differences in both the number of classes and frequency of each class in minicircle molecules from several kinetoplastid species [15,16,17,18,19,20,21,22,23]
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