ArticleParasites & vectors2017
The use of kDNA minicircle subclass relative abundance to differentiate between Leishmania (L.) infantum and Leishmania (L.) amazonensis.
Article in Parasites & vectors, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 23 papers.
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23 citing papers in PubMed, 51 citations in OpenAlex.
- Molecular Diagnosis of Leishmaniasis: Development of a qPCR Assay for Genus Detection and Differentiation ofDiagnostics (Basel, Switzerland) · 2026Article
- Sire risk factors for vertical transmission of Leishmania infantum by the dam.PLoS neglected tropical diseases · 2026Article
- Histone H3 K4 trimethylation occurs mainly at the origins of polycistronic transcription in the genome of Leishmania infantum promastigotes and intracellular amastigotes.BMC genomics · 2025Article
- Rapid monitoring of SARS-CoV-2 variants of concern through high-resolution melt analysis.Scientific reports · 2023Article
- High-resolution melting (HRM)-based detection of polymorphisms in the malic enzyme and glucose-6-phosphate isomerase genes for Leishmania infantum genotyping.Parasites & vectors · 2023Article
- Evaluation of Molecular Methods to Identify Chagas Disease and Leishmaniasis in Blood Donation Candidates in Two Brazilian Centers.Pathogens (Basel, Switzerland) · 2023Article
- Identification of a conserved maxicircle and unique minicircles as part of the mitochondrial genome of Leishmania martiniquensis strain PCM3 in Thailand.Parasites & vectors · 2022Article
- Deep kinetoplast genome analyses result in a novel molecular assay for detectingNAR genomics and bioinformatics · 2022Article
- Molecular Diagnosis of Leishmaniasis: Quantification of Parasite Load by a Real-Time PCR Assay with High Sensitivity.Pathogens (Basel, Switzerland) · 2021Article
- High resolution melting assay in discrimination of the main etiologic agents of leishmaniasis in Iran.Iranian journal of microbiology · 2021Article
- Efficacy of imiquimod 5% cream as first-line management in cutaneous leishmaniasis caused by Leishmania mexicana.Revista da Sociedade Brasileira de Medicina Tropical · 2021Article
- Evaluation of a kDNA-Based qPCR Assay for the Detection and Quantification of Old WorldMicroorganisms · 2020Article
- Performance of a universal PCR assay to identify different Leishmania species causative of Old World cutaneous leishmaniasis.Parasites & vectors · 2020Article
- Alkyl and Aryl Derivatives Based onMolecules (Basel, Switzerland) · 2020Article
- Differentiation ofMicroorganisms · 2020Article
- Data on the differentiation amongData in brief · 2020Article
- Exploring Leishmania infantum cathepsin as a new molecular marker for phylogenetic relationships and visceral leishmaniasis diagnosis.BMC infectious diseases · 2019Article
- Article
- Molecular Diagnosis of Visceral Leishmaniasis.Molecular diagnosis & therapy · 2018Review
- Real-time PCR applications for diagnosis of leishmaniasis.Parasites & vectors · 2018Review
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Authors and funding
8 authors at 3 institutions in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
backgroundLeishmaniasis is a neglected disease caused by many Leishmania species, belonging to subgenera Leishmania (Leishmania) and Leishmania (Viannia). Several qPCR-based molecular diagnostic approaches have been reported for detection and quantification of Leishmania species. Many of these approaches use the kinetoplast DNA (kDNA) minicircles as the target sequence. These assays had potential cross-species amplification, due to sequence similarity between Leishmania species. Previous works demonstrated discrimination between L. (Leishmania) and L. (Viannia) by SYBR green-based qPCR assays designed on kDNA, followed by melting or high-resolution melt (HRM) analysis. Importantly, these approaches cannot fully distinguish L. (L.) infantum from L. (L.) amazonensis, which can coexist in the same geographical area.
methodsDNA from 18 strains/isolates of L. (L.) infantum, L. (L.) amazonensis, L. (V.) braziliensis, L. (V.) panamensis, L. (V.) guyanensis, and 62 clinical samples from L. (L.) infantum-infected dogs were amplified by a previously developed qPCR (qPCR-ML) and subjected to HRM analysis; selected PCR products were sequenced using an ABI PRISM 310 Genetic Analyzer. Based on the obtained sequences, a new SYBR-green qPCR assay (qPCR-ama) intended to amplify a minicircle subclass more abundant in L. (L.) amazonensis was designed.
resultsThe qPCR-ML followed by HRM analysis did not allow discrimination between L. (L.) amazonensis and L. (L.) infantum in 53.4% of cases. Hence, the novel SYBR green-based qPCR (qPCR-ama) has been tested. This assay achieved a detection limit of 0.1 pg of parasite DNA in samples spiked with host DNA and did not show cross amplification with Trypanosoma cruzi or host DNA. Although the qPCR-ama also amplified L. (L.) infantum strains, the C
conclusionsA new and affordable SYBR-green qPCR-based approach to distinguish between L. (L.) infantum and L. (L.) amazonensis was developed exploiting the major abundance of a minicircle sequence rather than targeting a hypothetical species-specific sequence. The fast and accurate discrimination between these species can be useful to provide adequate prognosis and treatment.
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