Research
SYBR Green-based Real-Time PCR targeting kinetoplast DNA can be used to discriminate between the main etiologic agents of Brazilian cutaneous and visceral leishmaniases
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* Corresponding author: Constança Britto cbritto@ioc.fiocruz.br
1 Laboratório de Biologia Molecular e Doenças Endêmicas, Instituto Oswaldo Cruz, FIOCRUZ, Pavilhão Leônidas Deane, sala 209, Avenida Brasil 4365, Manguinhos, 21040-360 Rio de Janeiro, RJ, Brazil
2 Laboratório de Biologia Molecular de Insetos, Instituto Oswaldo Cruz, FIOCRUZ, Rio de Janeiro, RJ, Brazil
3 Laboratório Interdisciplinar em Pesquisas Médicas, Instituto Oswaldo Cruz, FIOCRUZ, Rio de Janeiro, RJ, Brazil
4 Laboratório de Bioquímica e Fisiologia de Insetos, Instituto Oswaldo Cruz, FIOCRUZ, Rio de Janeiro, RJ, Brazil
Parasites & Vectors 2012, 5:15 doi:10.1186/1756-3305-5-15
Published: 12 January 2012Additional files
Additional file 1:
Kinetoplast DNA minicircles conserved region sequences. This file contains the 12 generated sequences from amplified kDNA minicircles conserved region, representing three clones for each analyzed Leishmania reference strain [L. (V.) braziliensis, L. (V.) shawi, L. (L.) infantum chagasi, L. (L.) amazonensis].
Format: FAS Size: 2KB Download file
Additional file 2:
Melting curve analyses of the conserved motifs of kDNA amplicons. This file displays the characteristic SYBR Green dissociation profiles of kDNA amplified conserved regions after submitting the amplicons to a gradual temperature increase. The upper graphic represents reference strains of the Leishmania subgenus - L. amazonensis, L. infantum, with an estimated melting temperature (Tm) of 78.95°C ± 0.01. The lower graphic shows the resulting melting analysis for the Viannia subgenus reference strains - L. guyanensis, L. lainsoni, L. naiffi, L. braziliensis, L. shawi, where an average Tm of 77.36°C ± 0.02 was found. There was a significant difference between the Tm values of the two Leishmania subgenera (p < 0.001 - Mann-Whitney Rank Sum Test).
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