ArticleParasites & vectors2012
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.
Article in Parasites & vectors, 2012. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 39 papers.
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Who cites it
39 citing papers in PubMed.
- Molecular Diagnosis of Leishmaniasis: Development of a qPCR Assay for Genus Detection and Differentiation ofDiagnostics (Basel, Switzerland) · 2026Article
- Parasite Detection in Visceral Leishmaniasis Samples by Dye-Based qPCR Using New Gene Targets ofTropical medicine and infectious disease · 2023Article
- Evaluation of Molecular Methods to Identify Chagas Disease and Leishmaniasis in Blood Donation Candidates in Two Brazilian Centers.Pathogens (Basel, Switzerland) · 2023Article
- Socioeconomic Status of Guardians as a Risk Factor for Canine Visceral Leishmaniasis: A Cohort Study in an Endemic Area of the Federal District, Brazil.The American journal of tropical medicine and hygiene · 2023Article
- Detecting Leishmania in dogs: A hierarchical-modeling approach to investigate the performance of parasitological and qPCR-based diagnostic procedures.PLoS neglected tropical diseases · 2022Article
- Molecular Diagnosis of Leishmaniasis: Quantification of Parasite Load by a Real-Time PCR Assay with High Sensitivity.Pathogens (Basel, Switzerland) · 2021Article
- Diagnosis and Causative Species of Visceral Leishmaniasis in Southwest Saudi Arabia.The American journal of tropical medicine and hygiene · 2021Article
- High resolution melting assay in discrimination of the main etiologic agents of leishmaniasis in Iran.Iranian journal of microbiology · 2021Article
- Molecular Tracking of theFrontiers in cellular and infection microbiology · 2021Review
- Evaluation of a kDNA-Based qPCR Assay for the Detection and Quantification of Old WorldMicroorganisms · 2020Article
- One Health Approach to Leishmaniases: Understanding the Disease Dynamics through Diagnostic Tools.Pathogens (Basel, Switzerland) · 2020Review
- Differentiation ofMicroorganisms · 2020Article
- Trypanosomatid infections in captive wild mammals and potential vectors at the Brasilia Zoo, Federal District, Brazil.Veterinary medicine and science · 2020Article
- High resolution melting based method for rapid discriminatory diagnosis of co-infecting Leptomonas seymouri in Leishmania donovani-induced leishmaniasis.Parasitology international · 2020Article
- Impact of sequelae of visceral leishmaniasis and their contribution to ongoing transmission of Leishmania donovani.Pathogens and disease · 2019Review
- Natural infection by Leishmania infantum in the Lutzomyia longipalpis population of an endemic coastal area to visceral leishmaniasis in Brazil is not associated with bioclimatic factors.PLoS neglected tropical diseases · 2019Article
- Improving the reference standard for the diagnosis of canine visceral leishmaniasis: a challenge for current and future tests.Memorias do Instituto Oswaldo Cruz · 2019Article
- Evaluation of Real-time PCR for Diagnosis of Post-Kala-azar Dermal Leishmaniasis in Endemic Foci of Bangladesh.Open forum infectious diseases · 2018Article
- Molecular Diagnosis of Visceral Leishmaniasis.Molecular diagnosis & therapy · 2018Review
- Amino acid permease 3 (aap3) coding sequence as a target for Leishmania identification and diagnosis of leishmaniases using high resolution melting analysis.Parasites & vectors · 2018Article
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Abstract
backgroundLeishmaniases control has been hampered by the unavailability of rapid detection methods and the lack of suitable therapeutic and prophylactic measures. Accurate diagnosis, which can distinguish between Leishmania isolates, is essential for conducting appropriate prognosis, therapy and epidemiology. Molecular methods are currently being employed to detect Leishmania infection and categorize the parasites up to genus, complex or species level. Real-time PCR offers several advantages over traditional PCR, including faster processing time, higher sensitivity and decreased contamination risk.
resultsA SYBR Green real-time PCR targeting the conserved region of kinetoplast DNA minicircles was able to differentiate between Leishmania subgenera. A panel of reference strains representing subgenera Leishmania and Viannia was evaluated by the derivative dissociation curve analyses of the amplified fragment. Distinct values for the average melting temperature were observed, being 78.95 °C ± 0.01 and 77.36 °C ± 0.02 for Leishmania and Viannia, respectively (p < 0.05). Using the Neighbor-Joining method and Kimura 2-parameters, the alignment of 12 sequences from the amplified conserved minicircles segment grouped together L. (V.) braziliensis and L. (V.) shawii with a bootstrap value of 100%; while for L. (L.) infantum and L. (L.) amazonensis, two groups were formed with bootstrap values of 100% and 62%, respectively. The lower dissociation temperature observed for the subgenus Viannia amplicons could be due to a lower proportion of guanine/cytosine sites (43.6%) when compared to species from subgenus Leishmania (average of 48.4%). The method was validated with 30 clinical specimens from visceral or cutaneous leishmaniases patients living in Brazil and also with DNA samples from naturally infected Lutzomyia spp. captured in two Brazilian localities.
conclusionsFor all tested samples, a characteristic amplicon melting profile was evidenced for each Leishmania subgenus, corroborating the data from reference strains. Therefore, the analysis of thermal dissociation curves targeting the conserved kinetoplast DNA minicircles region is able to provide a rapid and reliable method to identify the main etiologic agents of cutaneous and visceral leishmaniases in endemic regions of Brazil.
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