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8. Domestic animals and epidemiology of visceral leishmaniasis, Nepal

Author

Bhattarai, Narayan Raj, Van der Auwera, Gert, Rijal, Suman, Picado, Albert, Speybroeck, Niko, Khanal, Basudha, De Doncker, Simonne, Das, Murari Lal, Ostyn, Bart, Davies, Clive, Coosemans, Marc, Berkvens, Dirk, Boelaert, Marleen, and Dujardin, Jean-Claude

Subjects
Geographic information systems -- Analysis, Goats -- Analysis, Infection -- Risk factors -- Analysis, Disease transmission -- Risk factors -- Analysis, Cattle -- Analysis, Epidemiology -- Analysis, Kala-azar -- Risk factors -- Analysis, Geographic information system, Health
Abstract

Visceral leishmaniasis (VL), also known as kala-azar, is a fatal vector-borne parasitic disease. Worldwide incidence is 500,000 cases per year; ≅ 90% of cases occur in India, Nepal, Bangladesh, Sudan, [...]

Published
2010
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14. Association of the Endobiont Double-Stranded RNA Virus LRV1 With Treatment Failure for Human Leishmaniasis Caused byLeishmania braziliensisin Peru and Bolivia

Author

Adaui, Vanessa, primary, Lye, Lon-Fye, additional, Akopyants, Natalia S., additional, Zimic, Mirko, additional, Llanos-Cuentas, Alejandro, additional, Garcia, Lineth, additional, Maes, Ilse, additional, De Doncker, Simonne, additional, Dobson, Deborah E., additional, Arevalo, Jorge, additional, Dujardin, Jean-Claude, additional, and Beverley, Stephen M., additional

Published
2015
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15. RFLP y RAPD para la tipificación de Leishmania neotropical

Author

Montalvo, Ana Margarita, Monzote, Lianet, Fraga, Jorge, Montano, Ivón, Muskus, Carlos, Marín, Marcel, de Doncker, Simonne, Vélez, Iván Darío, and Dujardin, Jean Claude

Subjects
Leishmania, polymerase chain reaction, polimorfismo de longitud del fragmento de restricción, restriction fragment length, leishmaniasis, reacción en cadena de la polimerasa, polymorphism
Abstract

Introducción. El análisis de la longitud de los fragmentos de restricción del producto amplificado y el estudio del ADN polimórfico amplificado al azar han demostrado ser herramientas útiles para la tipificación de Leishmania. Objetivos. Estudiar la utilidad de las técnicas moleculares para la identificación y tipificación de cepas de referencia de Leishmania spp. del Nuevo Mundo y valorar su aplicabilidad a muestras clínicas. Materiales y métodos. Se aplicó PCR para amplificar el gen que codifica la cisteíno-proteinasa B, y el análisis de la longitud de los fragmentos de restricción del producto amplificado utilizando ácido desoxirribonucleico de 16 cepas de referencia de Latinoamérica y de muestras clínicas de pacientes colombianos con leishmaniasis, y la técnica del ácido desoxirribonucleico polimórfico amplificado al azar utilizando ocho cepas de referencia. Se establecieron los patrones de bandas en cada caso. Resultados. Se obtuvo producto de amplificación en la PCR para Leishmania braziliensis, L. peruviana, L. panamensis y L. guyanensis. Para el resto, no fue posible amplificar el gen con los cebadores utilizados. La restricción mostró un patrón de bandas común para L. peruviana, L. guyanensis y L. panamensis, mientras L. braziliensis, presentaba un perfil individual único. El análisis de restricción del producto amplificado generó un patrón de bandas similar en los cinco pacientes estudiados, que se correspondía con el patrón generado por L. peruviana, L. guyanensis o L. panamensis. Mediante la amplificación al azar se obtuvieron patrones de bandas reproducibles con todas las cepas estudiadas, que posibilitaron la diferenciación. Se discuten las ventajas y limitaciones de ambos procederes. Conclusiones. El combinar ambas metodologías resultaría útil para identificar especies de importancia médica, tomando en cuenta sus ventajas y desventajas. Introduction. The analysis of the PCR-restriction fragment length polymorphism and random amplified polymorphic DNA have been useful tools for Leishmania identification. Objectives. Molecular procedures were demonstrated for identification and typing of reference strains of New World Leishmania and their applicability was validated for clinical samples. Materials and methods. DNA was extracted from 16 reference strains of Latin American Leishmania as well as from clinical samples of leishmaniasis patients. A sequence coding for cysteine proteinase B was amplified by PCR and subjected to restriction fragment length polymorphism analysis. The enzyme used was Taq1. For eight of the reference strains, the random amplified polymorphic desoxyribonucleic acid technique (RAPD) was applied. Band patterns for Leishmania species differentiation were established each each method. The sample size of the clinical sample was of 5. Results. PCR products of the cysteine proteinase B gene were obtained for L. braziliensis, L. peruviana, L. panamensis and L. guyanensis. For the other species, L. mexicana, L. amazonensis, L. garnhami, L. lainsoni, L. chagasi, L. naiffi, no amplification occurred. The patterns of restriction fragments revealed band patterns in common for L. peruviana, L. guyanensis and L. panamensis, whereas L. braziliensis had a distinctive pattern. When human samples were examined, amplification occurred for all cases, and the profiles corresponded to the common profile of L. peruviana, L. guyanensis and L. panamensis. The RAPD technique demonstrated reproducible and distinctive patterns for each of the 8 reference strains, L. mexicana, L. amazonensis, L. garnhami, L. lainsoni, L. chagasi, L. naiffi, making possible to differentiate all them. The advantages and limitations of each procedure are discussed. Conclusions. The combination of RFP and RAPD methodologies provide useful tools to identify medical important species of Leishmania by recognizing DNA sequences characteristic of each species.

Published
2008

16. Genome-wide SNP and microsatellite variation illuminate population-level epidemiology in the Leishmania donovani species complex

Author

Downing, Tim, primary, Stark, Olivia, additional, Vanaerschot, Manu, additional, Imamura, Hideo, additional, Sanders, Mandy, additional, Decuypere, Saskia, additional, de Doncker, Simonne, additional, Maes, Ilse, additional, Rijal, Suman, additional, Sundar, Shyam, additional, Dujardin, Jean-Claude, additional, Berriman, Matthew, additional, and Schönian, Gabriele, additional

Published
2012
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17. Multilocus genotyping reveals a polyphyletic pattern among naturally antimony-resistant Leishmania braziliensis isolates from Peru

Author

Adaui, Vanessa, primary, Maes, Ilse, additional, Huyse, Tine, additional, Van den Broeck, Frederik, additional, Talledo, Michael, additional, Kuhls, Katrin, additional, De Doncker, Simonne, additional, Maes, Louis, additional, Llanos-Cuentas, Alejandro, additional, Schönian, Gabriele, additional, Arevalo, Jorge, additional, and Dujardin, Jean-Claude, additional

Published
2011
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19. Whole genome sequencing of multiple Leishmania donovani clinical isolates provides insights into population structure and mechanisms of drug resistance

Author

Downing, Tim, primary, Imamura, Hideo, additional, Decuypere, Saskia, additional, Clark, Taane G., additional, Coombs, Graham H., additional, Cotton, James A., additional, Hilley, James D., additional, de Doncker, Simonne, additional, Maes, Ilse, additional, Mottram, Jeremy C., additional, Quail, Mike A., additional, Rijal, Suman, additional, Sanders, Mandy, additional, Schönian, Gabriele, additional, Stark, Olivia, additional, Sundar, Shyam, additional, Vanaerschot, Manu, additional, Hertz-Fowler, Christiane, additional, Dujardin, Jean-Claude, additional, and Berriman, Matthew, additional

Published
2011
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22. Comparative Gene Expression Analysis throughout the Life Cycle of Leishmania braziliensis: Diversity of Expression Profiles among Clinical Isolates

Author

Adaui, Vanessa, primary, Castillo, Denis, additional, Zimic, Mirko, additional, Gutierrez, Andres, additional, Decuypere, Saskia, additional, Vanaerschot, Manu, additional, De Doncker, Simonne, additional, Schnorbusch, Kathy, additional, Maes, Ilse, additional, Van der Auwera, Gert, additional, Maes, Louis, additional, Llanos-Cuentas, Alejandro, additional, Arevalo, Jorge, additional, and Dujardin, Jean-Claude, additional

Published
2011
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25. Immunological Determinants of Clinical Outcome in Peruvian Patients with Tegumentary Leishmaniasis Treated with Pentavalent Antimonials

Author

Maurer-Cecchini, Anne, primary, Decuypere, Saskia, additional, Chappuis, François, additional, Alexandrenne, Coralie, additional, De Doncker, Simonne, additional, Boelaert, Marleen, additional, Dujardin, Jean-Claude, additional, Loutan, Louis, additional, Dayer, Jean-Michel, additional, Tulliano, Gianfranco, additional, Arevalo, Jorge, additional, Llanos-Cuentas, Alexandro, additional, and Chizzolini, Carlo, additional

Published
2009
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26. PCR and direct agglutination asLeishmaniainfection markers among healthy Nepalese subjects living in areas endemic for Kala-Azar

Author

Bhattarai, Narayan Raj, primary, Van der Auwera, Gert, additional, Khanal, Basudha, additional, De Doncker, Simonne, additional, Rijal, Suman, additional, Das, Murari Lal, additional, Uranw, Surendra, additional, Ostyn, Bart, additional, Praet, Nicolas, additional, Speybroeck, Niko, additional, Picado, Albert, additional, Davies, Clive, additional, Boelaert, Marleen, additional, and Dujardin, Jean-Claude, additional

Published
2009
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27. Identification of Old World Leishmania spp. by specific polymerase chain reaction amplification of cysteine proteinase B genes and rapid dipstick detection

Author

Laurent, Thierry, primary, Van der Auwera, Gert, additional, Hide, Mallorie, additional, Mertens, Pascal, additional, Quispe-Tintaya, Wilber, additional, Deborggraeve, Stijn, additional, De Doncker, Simonne, additional, Leclipteux, Thierry, additional, Bañuls, Anne-Laure, additional, Büscher, Philippe, additional, and Dujardin, Jean-Claude, additional

Published
2009
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30. Epidemiological dynamics of antimonial resistance in Leishmania donovani: Genotyping reveals a polyclonal population structure among naturally-resistant clinical isolates from Nepal

Author

Laurent, Thierry, primary, Rijal, Suman, additional, Yardley, Vanessa, additional, Croft, Simon, additional, De Doncker, Simonne, additional, Decuypere, Saskia, additional, Khanal, Basudha, additional, Singh, Rupa, additional, Schönian, Gabriele, additional, and Kuhls, Katrin, additional

Published
2007
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37. Chromosome rearrangement in Leishmania mexicana M379

Author

Liu, Jianhua, primary, Gajendran, Nadesan, additional, Muthui, David, additional, Muyldermans, Serge, additional, Dujardin, Jean-Claude, additional, De Doncker, Simonne, additional, Jacquet, Diane, additional, Le Ray, Dominique, additional, Mathieu-Daudé, Françoise, additional, and Hamers, Raymond, additional

Published
1991
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38. Gene Expression Analysis of the Mechanism of Natural Sb(V) Resistance in Leishmania donovaniIsolates from Nepal

Author

Decuypere, Saskia, Rijal, Suman, Yardley, Vanessa, De Doncker, Simonne, Laurent, Thierry, Khanal, Basudha, Chappuis, François, and Dujardin, Jean-Claude

Abstract

ABSTRACTControl of visceral leishmaniasis (VL) is being challenged by the emergence of natural resistance against the first line of treatment, pentavalent antimonials [Sb(V)]. An insight into the mechanism of natural Sb(V) resistance is required for the development of efficient strategies to monitor the emergence and spreading of Sb(V) resistance in countries where VL is endemic. In this work, we have focused on the mechanism of natural Sb(V) resistance emerging in Nepal, a site where anthroponotic VL is endemic. Based on the current knowledge of Sb(V) metabolism and of the in vitro trivalent antimonial [Sb(III)] models of resistance to Leishmaniaspp., we selected nine genes for a comparative transcriptomic study on natural Sb(V)-resistant and -sensitive Leishmania donovaniisolates. Differential gene expression patterns were observed for the genes coding for 2-thiol biosynthetic enzymes, gamma-glutamylcysteine synthetase (GCS) and ornithine decarboxylase (ODC), and for the Sb(III) transport protein aquaglyceroporin 1 (AQP1). The results indicate that the mechanism for natural Sb(V) resistance partially differs from the mechanism reported for in vitro Sb(III) resistance. More specifically, we hypothesize that natural Sb(V) resistance results from (i) a changed thiol metabolism, possibly resulting in inhibition of Sb(V) activation in amastigotes, and (ii) decreased uptake of the active drug Sb(III) by amastigotes.

Published
2005
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39. Culture-Independent Species Typing of Neotropical Leishmaniafor Clinical Validation of a PCR-Based Assay Targeting Heat Shock Protein 70 Genes

Author

Garcia, Lineth, Kindt, Ann, Bermudez, Hernan, Llanos-Cuentas, Alejandro, De Doncker, Simonne, Arevalo, Jorge, Quispe Tintaya, Kelly Wilber, and Dujardin, Jean-Claude

Abstract

ABSTRACTPCR-restriction fragment length polymorphism analysis of heat shock protein 70 genes discriminates most neotropical Leishmaniaspecies, as well as Trypanosoma cruzi.The assay, combined with capillary electrophoresis in a microchip device, may be applied directly on clinical samples with a high sensitivity, hence supporting clinical and epidemiological monitoring of leishmaniasis.

Published
2004
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40. [PCR-RFLP and RAPD for typing neotropical Leishmania].

Author

Montalvo AM, Monzote L, Fraga J, Montano I, Muskus C, Marín M, de Doncker S, Vélez ID, and Dujardin JC

Subjects
Animals, Humans, Leishmaniasis etiology, Leishmaniasis physiopathology, Tropical Climate, Leishmania classification, Leishmania genetics, Polymerase Chain Reaction methods, Polymorphism, Restriction Fragment Length, Random Amplified Polymorphic DNA Technique
Abstract

Introduction: The analysis of the PCR-restriction fragment length polymorphism and random amplified polymorphic DNA have been useful tools for Leishmania identification., Objectives: Molecular procedures were demonstrated for identification and typing of reference strains of New World Leishmania and their applicability was validated for clinical samples., Materials and Methods: DNA was extracted from 16 reference strains of Latin American Leishmania as well as from clinical samples of leishmaniasis patients. A sequence coding for cysteine proteinase B was amplified by PCR and subjected to restriction fragment length polymorphism analysis. The enzyme used was Taq1. For eight of the reference strains, the random amplified polymorphic desoxyribonucleic acid technique (RAPD) was applied. Band patterns for Leishmania species differentiation were established each each method. The sample size of the clinical sample was of 5., Results: PCR products of the cysteine proteinase B gene were obtained for L. braziliensis, L. peruviana, L. panamensis and L. guyanensis. For the other species, L. mexicana, L. amazonensis, L. garnhami, L. lainsoni, L. chagasi, L. naiffi, no amplification occurred. The patterns of restriction fragments revealed band patterns in common for L. peruviana, L. guyanensis and L. panamensis, whereas L. braziliensis had a distinctive pattern. When human samples were examined, amplification occurred for all cases, and the profiles corresponded to the common profile of L. peruviana, L. guyanensis and L. panamensis. The RAPD technique demonstrated reproducible and distinctive patterns for each of the 8 reference strains, L. mexicana, L. amazonensis, L. garnhami, L. lainsoni, L. chagasi, L. naiffi, making possible to differentiate all them. The advantages and limitations of each procedure are discussed., Conclusions: The combination of RFP and RAPD methodologies provide useful tools to identify medical important species of Leishmania by recognizing DNA sequences characteristic of each species.

Published
2008

41. Direct identification of Leishmania species in biopsies from patients with American tegumentary leishmaniasis.

Author

Victoir K, De Doncker S, Cabrera L, Alvarez E, Arevalo J, Llanos-Cuentas A, Le Ray D, and Dujardin JC

Subjects
Animals, DNA Probes, DNA, Protozoan, Humans, Leishmania braziliensis classification, Leishmaniasis parasitology, Parasitology methods, Parasitology standards, Peru, Polymerase Chain Reaction methods, Polymorphism, Restriction Fragment Length, Sensitivity and Specificity, Leishmania braziliensis isolation & purification, Leishmaniasis diagnosis
Abstract

Accurate identification of Leishmania species is important for monitoring clinical outcome, adequately targeting treatment, and evaluation of epidemiological risk in tegumentary leishmaniasis. This is especially the case in regions where several species coexist and for travel medicine where the geographical source of infection is not always obvious. Species identification presently depends on parasite isolation, which is not very sensitive and not necessarily representative of parasites actually present in human tissues. We evaluated a polymerase chain reaction (PCR) assay combining amplification of the gp63 genes and restriction fragment length polymorphism (RFLP) analysis (gp63 PCR-RFLP) for direct Leishmania species-identification in tissues collected from Peruvian patients in 1999. By comparison with a kinetoplast DNA-based PCR, our PCR assay showed a detection sensitivity of 85%. Three species were encountered among patient samples, Leishmania (Viannia) braziliensis, L. (V.) peruviana and L. (V.) guyanensis, and their frequency and geographical distribution corresponded to earlier epidemiological studies of leishmaniasis in Peru. However, unexpected results raised questions about (i) the contribution of human migration to the emergence of new foci of given species, (ii) the pathogenicity of some species, and (iii) the frequency of mixed or hybrid infections.

Published
2003
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