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      Detección molecular de Leishmania (Viannia) lainsoni Silveira, Shaw, Braga & Ishikawa, 1987 (Kinetoplastida: Trypanosomatidae) en humanos de Paraguay Translated title: Detección molecular de Leishmania (Viannia) lainsoni Silveira, Shaw, Braga & Ishikawa, 1987 (Kinetoplastida: Trypanosomatidae) en humanos de Paraguay

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          Abstract

          RESUMEN La leishmaniasis es una de las enfermedades tropicales más desatendidas, causada por el parásito Leishmania. En Paraguay, la especie responsable de la leishmaniasis cutánea es (LC) es L. (Viannia) braziliensis. Aquí se reporta un caso diagnosticado de Leishmaniasis, y análisis moleculares empleando reacción en cadena de la polimerasa - polimorfismos de restricción de fragmentos de restricción (PCR-RFLP) demostraron que el caso fue causado por L. (V.) lainsoni. Este es el primer registro de esta especie para Paraguay, con lo cual se extiende el rango de distribución conocido de la especie, unos 1.430 km al sur de localidades previamente conocidas. Se necesitan más estudios para conocer la incidencia real de esta especie en casos de LC en Paraguay, y para identificar reservorios naturales del parásito en la naturaleza.

          Translated abstract

          ABSTRACT Leishmaniasis is one of the most neglected tropical diseases worldwide caused by the parasite Leishmania. In Paraguay the species responsible for cutaneous leishmaniasis (CL) is L. (Viannia) braziliensis. Here we report a case diagnosed with Leishmaniasis, and molecular analyses using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) demonstrate that the case was caused by L. (V.) lainsoni. This is the first record of this species for Paraguay, with which we extend the distribution range of the parasite 1,430 km southwards from the southernmost previous known locality. More studies are needed to know the actual incidence of this species in cases of CL in Paraguay, and to identify natural reservoirs in the wild.

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          Influence of Leishmania (Viannia) species on the response to antimonial treatment in patients with American tegumentary leishmaniasis.

          Pentavalent antimonials (SbV) are the first-line chemotherapy for American tegumentary leishmaniasis (ATL). There are, however, reports of the occurrence of treatment failure with these drugs. Few studies in Latin America have compared the response to SbV treatment in ATL caused by different Leishmania species. Clinical parameters and response to SbV chemotherapy were studied in 103 patients with cutaneous leishmaniasis (CL) in Peru. Leishmania isolates were collected before treatment and typed by multilocus polymerase-chain-reaction restriction fragment-length polymorphism analysis. The 103 isolates were identified as L. (Viannia) peruviana (47.6%), L. (V.) guyanensis (23.3%), L. (V.) braziliensis (22.3%), L. (V.) lainsoni (4.9%), L. (Leishmania) mexicana (1%), and a putative hybrid, L. (V.) braziliensis/L. (V.) peruviana (1%). L. (V.) guyanensis was most abundant in central Peru. Of patients infected with the 3 former species, 21 (21.9%) did not respond to SbV chemotherapy. The proportions of treatment failure (after 12 months of follow-up) were 30.4%, 24.5%, and 8.3% in patients infected with L. (V.) braziliensis, L. (V.) peruviana, and L. (V.) guyanensis, respectively. Infection with L. (V.) guyanensis was associated with significantly less treatment failure than L. (V.) braziliensis, as determined by multiple logistic regression analysis (odds ratio, 0.07 [95% confidence interval, 0.007-0.8]; P=.03). Leishmania species can influence SbV treatment outcome in patients with CL. Therefore, parasite identification is of utmost clinical importance, because it should lead to a species-oriented treatment.
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            Comparison of cutaneous leishmaniasis due to Leishmania (Viannia) braziliensis and L. (V.) guyanensis in Brazil: therapeutic response to meglumine antimoniate.

            We conducted a quasi-experimental study to compare the response to meglumine antimoniate in patients with localized cutaneous leishmaniasis from two endemic areas of Brazil that were infected by two Leishmania species. Sixty-one were infected by Leishmania (Viannia) braziliensis (group B) and 57 by L. (V.) guyanensis (group G). All had a parasitologically proven diagnosis and were treated with 20 mg of pentavalent antimonial (SbV)/kg/day given intravenously or intramuscularly for 20 days. Main outcomes were diagnosed using clinical criteria three months after treatment and patients were followed for six months. Intention-to-treat analysis showed a higher failure rate in group G (relative risk [RR] = 1.5, 95% confidence interval [CI] = 1.1-2.0, chi2 = 7.44, P = 0.006). The analysis using an explanatory approach including 52 patients from group B and 49 from group G, who were regularly treated and followed for six months, showed a low cure rate (50.8% in group B and 26.3% in group G) with a greater risk of failure in the latter group (RR = 1.7, 95% CI = 1.2-2.5, chi2 = 8.56, P = 0.003). The effect of the etiologic agent remained significant after adjusting for age, disease duration, and site and number of lesions that were identified as predictors of failure in a logistic regression model. We concluded that Leishmania species constitute an important factor in predicting the outcome of cutaneous leishmaniasis treated with a pentavalent antimonial.
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              Development and validation of PCR-based assays for diagnosis of American cutaneous leishmaniasis and identification of the parasite species.

              In this study, PCR assays targeting different Leishmania heat-shock protein 70 gene (hsp70) regions, producing fragments ranging in size from 230-390 bp were developed and evaluated to determine their potential as a tool for the specific molecular diagnosis of cutaneous leishmaniasis (CL). A total of 70 Leishmania strains were analysed, including seven reference strains (RS) and 63 previously typed strains. Analysis of the RS indicated a specific region of 234 bp in the hsp70 gene as a valid target that was highly sensitive for detection of Leishmania species DNA with capacity of distinguishing all analyzed species, after polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). This PCR assay was compared with other PCR targets used for the molecular diagnosis of leishmaniasis: hsp70 (1400-bp region), internal transcribed spacer (ITS)1 and glucose-6-phosphate dehydrogenase (G6pd). A good agreement among the methods was observed concerning the Leishmania species identification. Moreover, to evaluate the potential for molecular diagnosis, we compared the PCR targets hsp70-234 bp, ITS1, G6pd and mkDNA using a panel of 99 DNA samples from tissue fragments collected from patients with confirmed CL. Both PCR-hsp70-234 bp and PCR-ITS1 detected Leishmania DNA in more than 70% of the samples. However, using hsp70-234 bp PCR-RFLP, identification of all of the Leishmania species associated with CL in Brazil can be achieved employing a simpler and cheaper electrophoresis protocol.
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                Author and article information

                Contributors
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                Journal
                imt
                Revista del Instituto de Medicina Tropical
                Rev. Inst. Med. Trop.
                Instituto de Medicina Tropical (Asunción, , Paraguay )
                1996-3696
                June 2019
                : 14
                : 1
                : 21-28
                Affiliations
                [1] Asunción orgnameMinisterio de Salud Pública y Bienestar Social orgdiv1Programa Nacional Control Leishmaniasis (SENEPA) Paraguay
                [5] Asunción orgnameFundación ie, República de Colombia orgdiv1Laboratorio de Desarrollo Ambiental Paraguay
                [6] Asunción orgnameInstituto de Investigación Biológica del Paraguay Paraguay
                [7] Asunción orgnameOPS-OMS-Paraguay Paraguay
                [2] orgnameUniversidade de São Paulo orgdiv1Instituto de Biociências orgdiv2Laboratorio de Fisiologia de Tripanossomatídeos, Departamento de Fisiologia Brazil
                [4] São Paulo São Paulo orgnameUniversidade de São Paulo Brazil
                [3] Rio de Janeiro orgnamePan American Health Organization orgdiv1Communicable Diseases and Health Analysis Brazil
                Article
                S1996-36962019000100021
                10.18004/imt/201914121-28
                c9fcbc1e-7311-4999-aeb7-c43a74f3b8b6

                This work is licensed under a Creative Commons Attribution 4.0 International License.

                History
                : 14 April 2019
                : 01 April 2019
                Page count
                Figures: 0, Tables: 0, Equations: 0, References: 28, Pages: 8
                Product

                SciELO Paraguay

                Categories
                Articulos Originales

                South America,Epidemiología,Leishmaniasis,PCR-RFLP,Sudamérica.,Epidemiology

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