Simplified molecular detection of Leishmania parasites in various clinical samples from patients with leishmaniasis

Mugasa, Claire M.; Laurent, Thierry; Schoone, Gerard J.; Basiye, Frank; Saad, Alfarazdeg A.; Safi, Sayda El; Kager, Piet A.; Schallig, Henk D. F. H.

doi:10.1186/1756-3305-3-13

Cited by 38 publications

(35 citation statements)

References 25 publications

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“…Thus, correct diagnosis of the infecting Leishmania species is crucial for making decisions regarding prognosis and treatments, and also for epidemiological monitoring of the parasite spread [4]. …”

Section: Introductionmentioning

confidence: 99%

Sequence analysis of the 3’-untranslated region of HSP70 (type I) genes in the genus Leishmania: its usefulness as a molecular marker for species identification

et al. 2012

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BackgroundThe Leishmaniases are a group of clinically diverse diseases caused by parasites of the genus Leishmania. To distinguish between species is crucial for correct diagnosis and prognosis as well as for treatment decisions. Recently, sequencing of the HSP70 coding region has been applied in phylogenetic studies and for identifying of Leishmania species with excellent results.MethodsIn the present study, we analyzed the 3’-untranslated region (UTR) of Leishmania HSP70-type I gene from 24 strains representing eleven Leishmania species in the belief that this non-coding region would have a better discriminatory capacity for species typing than coding regions.ResultsIt was observed that there was a remarkable degree of sequence conservation in this region, even between species of the subgenus Leishmania and Viannia. In addition, the presence of many microsatellites was a common feature of the 3´-UTR of HSP70-I genes in the Leishmania genus. Finally, we constructed dendrograms based on global sequence alignments of the analyzed Leishmania species and strains, the results indicated that this particular region of HSP70 genes might be useful for species (or species complex) typing, improving for particular species the discrimination capacity of phylogenetic trees based on HSP70 coding sequences. Given the large size variation of the analyzed region between the Leishmania and Viannia subgenera, direct visualization of the PCR amplification product would allow discrimination between subgenera, and a HaeIII-PCR-RFLP analysis might be used for differentiating some species within each subgenera.ConclusionsSequence and phylogenetic analyses indicated that this region, which is readily amplified using a single pair of primers from both Old and New World Leishmania species, might be useful as a molecular marker for species discrimination.

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Section: Introductionmentioning

confidence: 99%

Sequence analysis of the 3’-untranslated region of HSP70 (type I) genes in the genus Leishmania: its usefulness as a molecular marker for species identification

et al. 2012

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“…Our AuNP/MB based electrochemical approach results are better than those obtained by other POC tests using nucleic acid sequence based amplification and coupled to oligochromatography for Leishmania detection, and even considerably better than the one parasite per PCR detection limit offered by the OligoC‐test…”

Section: Resultsmentioning

confidence: 67%

Magnetic Bead/Gold Nanoparticle Double-Labeled Primers for Electrochemical Detection of Isothermal AmplifiedLeishmaniaDNA

Escosura‐Muñiz

Baptista-Pires

Serrano³

et al. 2015

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A novel methodology for the isothermal amplification of Leishmania DNA using labeled primers combined with the advantages of magnetic purification/preconcentration and the use of gold nanoparticle (AuNP) tags for the sensitive electrochemical detection of such amplified DNA is developed. Primers labeled with AuNPs and magnetic beads (MBs) are used for the first time for the isothermal amplification reaction, being the amplified product ready for the electrochemical detection. The electrocatalytic activity of the AuNP tags toward the hydrogen evolution reaction allows the rapid quantification of the DNA on screen-printed carbon electrodes. Amplified products from the blood of dogs with Leishmania (positive samples) are discriminated from those of healthy dogs (blank samples). Quantitative studies demonstrate that the optimized method allows us to detect less than one parasite per microliter of blood (8 × 10(-3) parasites in the isothermal amplification reaction). This pioneering approach is much more sensitive than traditional methods based on real-time polymerase chain reaction (PCR), and is also more rapid, cheap, and user-friendly.

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“…Although such assays are more sensitive than conventional PCR, their high costs make this test not appropriate in a field setting. Another form of PCR such as OligoC-test [100], PCR-ELISA [101] and Nucleic acid sequence based amplification (NASBA) have been developed and found to be more sensitive than conventional PCR [43]. More recently, rapid and highly specific loop mediated isothermal amplification (LAMP) has emerged as powerful tool for point of care diagnosis and has been validated on VL & PKDL in several countries [102, 103].…”

Section: Standard Diagnostic Tools and Their Limitationsmentioning

confidence: 99%

Molecular Diagnosis of Visceral Leishmaniasis

Sundar

2018

Mol Diagn Ther

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Visceral leishmaniasis (VL), a deadly parasitic disease, is a major public health concern globally. Countries affected by VL have signed the London Declaration on Neglected Tropical Diseases and committed to eliminate VL as a public health problem by 2020. To achieve and sustain VL elimination, it will become progressively important not to miss any remaining cases in the community who can maintain transmission. This requires accurate identification of symptomatic and asymptomatic carriers using highly sensitive diagnostic tools at the primary health service setting. The rK39 rapid diagnostic test (RDT) is the most widely used tool and with its good sensitivity and specificity is the first choice for decentralized diagnosis of VL in endemic areas. However, this test cannot discriminate between current, subclinical, or past infections and is useless for diagnosis of relapses and as a prognostic (cure) test. Importantly, as the goal of elimination of VL as a public health problem is approaching, the number of people susceptible to infection will increase. Therefore, correct diagnosis using a highly sensitive diagnostic test is crucial for applying appropriate treatment and management of cases. Recent advances in molecular techniques have improved Leishmania detection and quantification, and therefore this technology has become increasingly relevant due to its possible application in a variety of clinical sample types. Most importantly, given current problems in identifying asymptomatic individuals because of poor correlation between the main methods of detection, molecular tests are valuable for VL elimination programs, especially to monitor changes in burden of infection in specific communities. This review provides a comprehensive overview of the available VL diagnostics and discusses the usefulness of molecular methods in the diagnosis, quantification, and species differentiation as well as their clinical applications.

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Simplified molecular detection of Leishmania parasites in various clinical samples from patients with leishmaniasis

Cited by 38 publications

References 25 publications

Sequence analysis of the 3’-untranslated region of HSP70 (type I) genes in the genus Leishmania: its usefulness as a molecular marker for species identification

Sequence analysis of the 3’-untranslated region of HSP70 (type I) genes in the genus Leishmania: its usefulness as a molecular marker for species identification

Magnetic Bead/Gold Nanoparticle Double-Labeled Primers for Electrochemical Detection of Isothermal AmplifiedLeishmaniaDNA

Molecular Diagnosis of Visceral Leishmaniasis

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