ArticleParasites & vectors2016
Real-time PCR using FRET technology for Old World cutaneous leishmaniasis species differentiation.
Article in Parasites & vectors, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 13 papers, 2 of them syntheses that pooled it.
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Who cites it
13 citing papers in PubMed, 2 syntheses or guidelines pooled it, 31 citations in OpenAlex.
- Diagnostic performance of CL Detect rapid-immunochromatographic test for cutaneous leishmaniasis: a systematic review and meta-analysis.Systematic reviews · 2023Pooled it
- Diagnosing Cutaneous leishmaniasis using FluorescencePathogens and global health · 2019Pooled it
- From Skin to Sequence: Pan-species Detection and Speciation of Leishmania.The Journal of infectious diseases · 2026Article
- Mouse Models for Use in Cryptosporidium Infection Studies, Quantification of Parasite Burden Using Flow Cytometry, qPCR and Histopathology, and Confocal Imaging of Oocysts.Methods in molecular biology (Clifton, N.J.) · 2026Article
- Advances and Challenges in the Diagnosis of Leishmaniasis.Molecular diagnosis & therapy · 2025Review
- Sampling is decisive to determination of Leishmania (Viannia) species.PLoS neglected tropical diseases · 2024Article
- Recombinase-based amplification coupled with lateral flow chromatography for the specific and sensitive detection and identification ofFrontiers in microbiology · 2024Article
- FRET Based Biosensor: Principle Applications Recent Advances and Challenges.Diagnostics (Basel, Switzerland) · 2023Review
- Laboratory diagnostics for human Leishmania infections: a polymerase chain reaction-focussed review of detection and identification methods.Parasites & vectors · 2022Review
- Establish an allele-specific real-time PCR for Leishmania species identification.Infectious diseases of poverty · 2022Article
- Identification ofJournal of parasitic diseases : official organ of the Indian Society for Parasitology · 2021Article
- Prevalence of and Factors Associated with Negative Microscopic Diagnosis of Cutaneous Leishmaniasis in Rural Peru.The American journal of tropical medicine and hygiene · 2018Article
- Real-time PCR applications for diagnosis of leishmaniasis.Parasites & vectors · 2018Review
Corrections and comments
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Authors and funding
8 authors at 5 institutions in 3 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
backgroundRecently, there has been a re-emergence of cutaneous leishmaniasis in endemic countries and an increase in imported cases in non-endemic countries by travelers, workers, expatriates, immigrants, and military force personnel. Old World cutaneous leishmaniasis is caused primarily by Leishmania major, L. tropica and L. aethiopica. Despite their low sensitivity, diagnosis traditionally includes microscopic and histopathological examinations, and in vitro cultivation. Several conventional PCR techniques have been developed for species identification, which are time-consuming and labour-intensive. Real-time PCR using SYBR green dye, although provides rapid detection, may generate false positive signals. Therefore, a rapid and easy method such as a FRET-based real-time PCR would improve not only the turn-around time of diagnosing Old World cutaneous Leishmania species but will also increase its specificity and sensitivity.
methodsA FRET-based real-time PCR assay which amplifies the cathepsin L-like cysteine protease B gene encoding a major Leishmania antigen was developed to differentiate L. major, L. tropica, and L. aethiopica in one single step using one set of primers and probes. Assay performance was tested on cutaneous and visceral strains of Leishmania parasite cultures and isolates of other protozoan parasites as well as human biopsy specimen.
resultsThe assay readily differentiates between the three Old World cutaneous leishmaniasis species based on their melting curve characteristics. A single Tm at 55.2 ± 0.5 °C for L. aethiopica strains was distinguished from a single Tm at 57.4 ± 0.2 °C for L. major strains. A double curve with melting peaks at 66.6 ± 0.1 °C and 48.1 ± 0.5 °C or 55.8 ± 0.6 °C was observed for all L. tropica strains. The assay was further tested on biopsy specimens, which showed 100% agreement with results obtained from isoenzyme electrophoresis and Sanger sequencing.
conclusionCurrently, there are no published data on real-time PCR using FRET technology to differentiate between Old World cutaneous Leishmania species. In summary, our assay based on specific hybridization addresses the limitations of previous PCR technology and provides a single step, reliable method of species identification and rapid diagnostic applications.
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