Sequence analysis and PCR-RFLP profiling of the hsp70 gene as a valuable tool for identifying Leishmania species associated with human leishmaniasis in Brazil

Silva, Leonardo Alves da; Sousa, Cíntia dos Santos de; Graça, Grazielle Cardoso da; Porrozzi, Renato; Cupolillo, Elisa

doi:10.1016/j.meegid.2009.11.001

Cited by 81 publications

(92 citation statements)

References 38 publications

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“…However, the kDNA-PCR method was positive in 19 patients, indicating that L. (V.) braziliensis is the predominant species, although similar electrophoretic patterns may occur within related species from Viannia subgenus, as some studies have reported. [17][18][19] Other species can be important in specific geographical situations, and molecular methods play a pivotal role in defining different clinical presentations, response to treatment, and choice of the best drug for each case. Guerra and others reported 46 cases of ML along the Amazon region, with 16 cases caused by L. (V.) guyanensis, thus showing that this species is also a significant causative agent within the region.…”

Section: Discussionmentioning

confidence: 99%

Efficacy and Safety of Liposomal Amphotericin B for the Treatment of Mucosal Leishmaniasis from the New World: A Retrospective Study

Cunha¹,

Leão²,

Soler³

et al. 2015

The American Society of Tropical Medicine and Hygiene

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Abstract. The standard treatment of mucosal leishmaniasis (ML) is pentavalent antimonials, agents with serious adverse effects. Alternative agents include amphotericin B deoxycholate and liposomal amphotericin B. We performed a retrospective study including 29 patients treated with liposomal amphotericin B, most of whom had comorbidities, history of previous treatment of ML, and contraindications to the use of antimonial pentavalent or amphotericin B deoxycholate. We observed a cure rate of 93.1%. Kidney failure was the most important side effect, reported in five patients (17.2%). This study showed a good efficacy and safety profile of liposomal amphotericin B in patients with ML and contraindications to the use of other agents.

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Section: Discussionmentioning

confidence: 99%

Efficacy and Safety of Liposomal Amphotericin B for the Treatment of Mucosal Leishmaniasis from the New World: A Retrospective Study

Cunha¹,

Leão²,

Soler³

et al. 2015

The American Society of Tropical Medicine and Hygiene

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“…The restriction endonuclease HaeIII produces diagnostic RFLP patterns for the L. guyanensis species complex as well as for L. lainsoni and L. shawi (Garcia et al 2004;Da Silva et al 2010). However, L. braziliensis and L. peruviana, both belonging to the L. braziliensis complex, as well as L. naiffi share an identical HaeIII RFLP pattern and can only be distingished by using a second restriction enzyme (Da Silva et al 2010). The same is true for L. guyanensis and L. panamensis both belonging to the L. guyanensis complex (Fraga et al 2010).…”

Section: O L E C U L a R E P I D E M I O L O G Y A N D M O L E C U mentioning

confidence: 99%

Molecular approaches for a better understanding of the epidemiology and population genetics ofLeishmania

2010

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S U M M A R YMolecular approaches are being used increasingly for epidemiological studies of visceral and cutaneous leishmaniases. Several molecular markers resolving genetic differences between Leishmania parasites at species and strain levels have been developed to address key epidemiological and population genetic questions. The current gold standard, multilocus enzyme typing (MLEE), needs cultured parasites and lacks discriminatory power. PCR assays identifying species directly with clinical samples have proven useful in numerous field studies. Multilocus sequence typing (MLST) is potentially the most powerful phylogenetic approach and will, most probably, replace MLEE in the future. Multilocus microsatellite typing (MLMT) is able to discriminate below the zymodeme level and seems to be the best candidate for becoming the gold standard for distinction of strains. Population genetic studies by MLMT revealed geographical and hierarchic population structure in L. tropica, L. major and the L. donovani complex. The existence of hybrids and gene flow between Leishmania populations suggests that sexual recombination is more frequent than previously thought. However, typing and analytical tools need to be further improved. Accessible databases should be created and sustained for integrating data obtained by different researchers. This would allow for global analyses and help to avoid biases in analyses due to small sample sizes.

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“…Incluso, se confirmó la enfermedad en un paciente tras 40 años de haber abandonado el área endémica (25), lo cual evidencia su importancia. (23). Más recientemente se han reportado algoritmos que permiten esta discriminación mediante la amplificación y la posterior digestión de fragmentos más cortos del mismo gen hsp70, lo que se validó utilizando ADN de cuatro cepas de referencia (27).…”

Section: Discussionunclassified

Identificación molecular de aislamientos clínicos de Leishmania spp. procedentes de Colombia con base en el gen hsp70

et al. 2016

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Introducción. La leishmaniasis es una enfermedad de alta prevalencia en Colombia, donde al menos seis especies diferentes pueden causar una variada presentación clínica en el humano. La tipificación de la especie es importante no solo desde el punto de vista epidemiológico, sino en el diagnóstico, dado que el tratamiento y el esquema de tratamiento pueden variar dependiendo de la especie identificada. Para la identificación se han utilizado distintas alternativas metodológicas, con variable poder discriminatorio.Objetivo. Realizar la identificación molecular de especies de Leishmania spp. mediante la amplificación molecular de un fragmento del gen hsp70.Materiales y métodos. Se realizó la amplificación molecular de un fragmento del gen hsp70: PCR-hsp70 (siglas en inglés) seguida del análisis del tamaño de los fragmentos de restricción (RFLP siglas en inglés), a 81 aislamientos clínicos de Leishmania spp., provenientes de pacientes con enfermedad cutánea y mucocutánea, en los cuales se identificaron las especies presentes.Resultados. Se obtuvo un único producto de amplificación para el total de muestras analizadas. La restricción enzimática permitió identificar 70 aislamientos con un patrón de bandas correspondiente a Leishmania braziliensis, que incluye dos patrones diferentes (62 y 8 aislamientos respectivamente); 9 aislamientos compatibles con Leishmania panamensis y 2 con Leishmania guyanensis. El origen geográfico de los aislamientos concuerda con reportes anteriores sobre la distribución de las especies correspondientes.Conclusiones. La técnica de PCR-hsp70/RFLP utilizada es útil para identificar especies de Leishmania aisladas de muestras clínicas de Colombia que puede ser aplicable también al estudio de cepas provenientes de vectores y reservorios con importancia epidemiológica.

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Sequence analysis and PCR-RFLP profiling of the hsp70 gene as a valuable tool for identifying Leishmania species associated with human leishmaniasis in Brazil

Cited by 81 publications

References 38 publications

Efficacy and Safety of Liposomal Amphotericin B for the Treatment of Mucosal Leishmaniasis from the New World: A Retrospective Study

Efficacy and Safety of Liposomal Amphotericin B for the Treatment of Mucosal Leishmaniasis from the New World: A Retrospective Study

Molecular approaches for a better understanding of the epidemiology and population genetics ofLeishmania

Identificación molecular de aislamientos clínicos de Leishmania spp. procedentes de Colombia con base en el gen hsp70

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