Molecular Diagnosis of Leishmaniasis: Current Status and Future Applications

Reithinger, Richard; Dujardin, Jean‐Claude

doi:10.1128/jcm.02029-06

Cited by 354 publications

(294 citation statements)

References 28 publications

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“…14 , 15 While these techniques have increased sensitivity, can detect active infections, and can be used for CL patients; they also require sufficient technical skill, equipment, continual electricity, and are considerably more expensive than the serological tests. 16 Therefore, many of these tests are not suitable for diagnosis in endemic regions, except in wellequipped laboratories. Isothermal amplification techniques such as nucleic acid sequence-based amplification (NASBA) for leishmaniasis have also been developed 17 ; however, the post-amplification analysis with oligo-chromatography makes this technique time-consuming and contamination prone.…”

Section: Introductionmentioning

confidence: 99%

Development of a Reverse Transcriptase Loop-Mediated Isothermal Amplification (LAMP) Assay for the Sensitive Detection of Leishmania Parasites in Clinical Samples

Adams¹,

Schoone²,

Ageed³

et al. 2010

The American Journal of Tropical Medicine and Hygiene

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Abstract. Here we describe a generic, reverse transcriptase-loop-mediated isothermal amplification (RT-LAMP) assay, for the identification of Leishmania species from clinical samples. LAMP is an isothermal reaction recently developed as a point-of-care diagnostic tool. Primers were designed in the conserved region of the 18S ribosomal RNA (rRNA) gene; amplification was visualized by the pre-amplification addition of fluorescent detection reagent (FDR) and a simple UV lamp. By using a reverse-transcriptase step, the system detected infections between 10 and 100 parasites per mL. The assay was tested on a range of nucleic acid extracts from Leishmania species, visceral leishmaniasis (VL) patients from Sudan, and cutaneous leishmaniasis (CL) patients from Suriname. The sensitivity of RT-LAMP from the blood of VL patients was 83% ( N = 30) compared with microscopy of bone-marrow and lymph-node aspirates; for CL patients the observed sensitivity was 98% ( N = 43). The potential to use LAMP as a diagnostic tool for leishmaniasis is discussed.

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Section: Introductionmentioning

confidence: 99%

Development of a Reverse Transcriptase Loop-Mediated Isothermal Amplification (LAMP) Assay for the Sensitive Detection of Leishmania Parasites in Clinical Samples

Adams¹,

Schoone²,

Ageed³

et al. 2010

The American Journal of Tropical Medicine and Hygiene

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“…Rapid and more efficient serological tests have been developed and validated as a tool to supplement the microscopic diagnosis. 6,7 Immunoserologic studies for VL in dogs, such as direct agglutination test (DAT), 8 indirect immunofluorescence assay (IFA), 4 enzyme-linked immunosorbent assay (ELISA), 6,7 polymerase chain reaction, 9 and immunochromatographic tests 6,7 (ICTs) were adapted from assays validated in humans, [10][11][12][13] with different levels of performance in canine VL. A recombinant fusion protein with tandem repeats of k39 kinesin regions and K26/HASPb1 when used in ELISA was found to detect high levels of antibody responses in infected patients.…”

Section: Introductionmentioning

confidence: 99%

An rK28-Based Immunoenzymatic Assay for the Diagnosis of Canine Visceral Leishmaniasis in Latin America

Lauricella

Maidana

Frias

et al. 2016

The American Society of Tropical Medicine and Hygiene

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Abstract. Direct observation of Leishmania parasites in tissue aspirates has shown low sensitivity for the detection of canine visceral leishmaniasis (VL). Therefore in the last quarter century immunoenzymatic tests have been developed to improve diagnosis. The purpose of this study was to develop a fast recombinant K28 antigen, naked-eye qualitative enzyme-linked immunosorbent assay (VL Ql-ELISA) and a quantitative version (VL Qt-ELISA), and to display it in a kit format, whose cutoff value (0.156) was selected as the most adequate one to differentiate reactive from nonreactive samples. Considering 167 cases and 300 controls, sensitivity was 91% for both assays and specificity was 100% and 98.7% in Ql-ELISA and Qt-ELISA, respectively. Positive predictive values were 100% and 97.4% for Ql-ELISA and Qt-ELISA, respectively, and negative predictive values were 95.2% for both ELISAs. Reagent stability, reliability studies, including periodic repetitions and retest of samples, cutoff selection, and comparison of rK28 ELISAs with rK39 immunochromatographic test, were the international criteria that supported the quality in both kits. The performance of both ELISA kits in this work confirmed their validity and emphasized their usefulness for low-to-medium complexity laboratories.

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“…[1][2][3][4] An estimated 1.5 to 2 million new cases of leishmaniasis occurs each year in the world, which in the visceral manifestation is often fatal if untreated. [5][6][7][8][9] Unfortunately non-availability of satisfactory chemotherapeutic agents and failure to develop an effective Synthesis, X-ray Crystal Structure and Theoretical Calculations J. Braz. Chem.…”

Section: Introductionmentioning

confidence: 99%

Synthesis, X-ray crystal structure and theoretical calculations of antileishmanial neolignan analogues

Nascimento¹,

Santos²,

Santos³

et al. 2010

J. Braz. Chem. Soc.

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A síntese e a estrutura cristalina por difração de raios-X de dois análogos de neolignanas, 2-(4-clorofenil)-1-feniletanona (20) e 2-[tio(4-clorofenil)]-1-(3,4-dimetoxifenil)propan-1-ona (12) são descritas. O composto 12 apresenta atividade intracelular contra Leishmania donovani e Leishmania amazonensis de amastigotas que causam a leishmaniose tegumentar e visceral. Além disso, a teoria do funcional de densidade (DFT) com o funcional híbrido B3LYP foi empregado para calcular um conjunto de descritores moleculares para dezenove análogos sintéticos de neolignanas com atividades antileishmaniose. Posteriormente, a análise discriminante stepwise foi realizada para investigar possíveis relações entre a estrutura molecular e atividades biológicas. Por meio dessa análise os compostos foram classificados em dois grupos ativos e inativos de acordo com seu grau de atividade biológica, e as propriedades mais importantes foram as cargas de alguns átomos, a afinidade eletrônica e o ClogP.The synthesis and X-ray crystal diffraction structure of two analogues of neolignans, 2-(4-chlorophenyl)-1-phenylethanone (20) and 2-[(4-chlorophenyl)thio]-1-(3,4-dimethoxyphenyl) propan-1-one (12) is described. The compound 12 presents activity against intracellular Leishmania donovani and Leishmania amazonensis amastigotes that cause cutaneous and visceral leishmaniasis. In addition, the density functional theory (DFT) with the B3LYP hybrid functional was employed to calculate a set of molecular descriptors for nineteen synthetic analogues of neolignans with antileishmanial activities. Afterwards, the stepwise discriminant analysis was performed to investigate possible relationship between the molecular descriptors and biological activities. Through this analysis the compounds were classified into two groups active and inactive according to their degree of biological activities, and the more important properties were charges on some key atoms, electronic affinity and ClogP.

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Molecular Diagnosis of Leishmaniasis: Current Status and Future Applications

Cited by 354 publications

References 28 publications

Development of a Reverse Transcriptase Loop-Mediated Isothermal Amplification (LAMP) Assay for the Sensitive Detection of Leishmania Parasites in Clinical Samples

Development of a Reverse Transcriptase Loop-Mediated Isothermal Amplification (LAMP) Assay for the Sensitive Detection of Leishmania Parasites in Clinical Samples

An rK28-Based Immunoenzymatic Assay for the Diagnosis of Canine Visceral Leishmaniasis in Latin America

Synthesis, X-ray crystal structure and theoretical calculations of antileishmanial neolignan analogues

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