<?xml version="1.0" encoding="ISO-8859-1"?><article xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance">
<front>
<journal-meta>
<journal-id>1812-9528</journal-id>
<journal-title><![CDATA[Memorias del Instituto de Investigaciones en Ciencias de la Salud]]></journal-title>
<abbrev-journal-title><![CDATA[Mem. Inst. Investig. Cienc. Salud]]></abbrev-journal-title>
<issn>1812-9528</issn>
<publisher>
<publisher-name><![CDATA[Instituto de Investigaciones en Ciencias de la Salud]]></publisher-name>
</publisher>
</journal-meta>
<article-meta>
<article-id>S1812-95282016000300024</article-id>
<article-id pub-id-type="doi">10.18004/mem.iics/1812-9528/2016.014(03)24-033</article-id>
<title-group>
<article-title xml:lang="es"><![CDATA[Estandarización de la técnica de PCR-RFLP de la región ITS1, para la caracterización molecular de Leishmania (L.) infantum, en muestras caninas]]></article-title>
<article-title xml:lang="en"><![CDATA[Standardization of PCR-RFLP technique of the ITS-1 region for Leishmania (L.) infantum molecular characterization in canine samples]]></article-title>
</title-group>
<contrib-group>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Chaves]]></surname>
<given-names><![CDATA[Enmanuel Céspedes]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Portillo]]></surname>
<given-names><![CDATA[Nilda]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Thomaz-Soccol]]></surname>
<given-names><![CDATA[Vanette]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Gonçalves]]></surname>
<given-names><![CDATA[André Luiz]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Brítez]]></surname>
<given-names><![CDATA[Nilsa González]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
</contrib-group>
<aff id="Af1">
<institution><![CDATA[,Universidad Nacional de Asunción Instituto de Investigaciones en Ciencias de la Salud Departamento de Medicina Tropical]]></institution>
<addr-line><![CDATA[ ]]></addr-line>
<country>Paraguay</country>
</aff>
<aff id="Af2">
<institution><![CDATA[,Universidad Federal del Paraná Laboratorio de Biología Molecular del Programa de Post-graduación en Ingeniería de Bioprocesos y Biotecnología ]]></institution>
<addr-line><![CDATA[ ]]></addr-line>
<country>Brasil</country>
</aff>
<pub-date pub-type="pub">
<day>00</day>
<month>12</month>
<year>2016</year>
</pub-date>
<pub-date pub-type="epub">
<day>00</day>
<month>12</month>
<year>2016</year>
</pub-date>
<volume>14</volume>
<numero>3</numero>
<fpage>24</fpage>
<lpage>33</lpage>
<copyright-statement/>
<copyright-year/>
<self-uri xlink:href="http://scielo.iics.una.py/scielo.php?script=sci_arttext&amp;pid=S1812-95282016000300024&amp;lng=en&amp;nrm=iso"></self-uri><self-uri xlink:href="http://scielo.iics.una.py/scielo.php?script=sci_abstract&amp;pid=S1812-95282016000300024&amp;lng=en&amp;nrm=iso"></self-uri><self-uri xlink:href="http://scielo.iics.una.py/scielo.php?script=sci_pdf&amp;pid=S1812-95282016000300024&amp;lng=en&amp;nrm=iso"></self-uri><abstract abstract-type="short" xml:lang="es"><p><![CDATA[RESUMEN La leishmaniasis es una enfermedad producida por protozoarios parásitos del género Leishmania. Estos parásitos infectan a hospedadores mamíferos, entre los cuales los perros han sido implicados como reservorios del parásito. Este trabajo planteó estandarizar la técnica de la PCR-RFLP luego de la amplificación de la región ITS1 de Leishmania spp, como herramienta útil para la detección y caracterización molecular. Se utilizaron promastigotes de cultivo y muestras de biopsias procedentes de perros con leishmaniasis visceral previamente diagnosticados en el Centro Antirrábico Nacional. La región ITS1 del ADN genómico nuclear de Leishmania spp. fue amplificada utilizando los cebadores LITSR y L5,8S. La técnica ITS1 PCR-RFLP aplicada, permitió la detección de Leishmania (L) infantum en 10/10 aislados de parásitos mantenidos en medio NNN, en 10/18 muestras de bazo y 10/18 muestras de ganglio linfático poplíteo. Las condiciones óptimas de reacción fueron 0,2 mM de dNTPs, 0,1 pmol de cada cebador y 1U de Taq polimerasa. La sensibilidad de la PCR fue de 3 ng/µL de ADN en aislados de cultivo NNN y 60 ng/µL de ADN en muestras de biopsias, mientras que la especificidad fue de 100% para la detección de Leishmania sp. La enzima de restricción Hae III, determinó fragmentos de 184, 72 y 55 pb., que resultaron específicos para la especie Leishmania (L.) infantum. El marcador utilizado resultó confiable para la detección y caracterización de Leishmania sp. en perros procedentes de zonas endémicas, lo cual podría ser útil para verificar las especies de parásitos circulantes entre los perros.]]></p></abstract>
<abstract abstract-type="short" xml:lang="en"><p><![CDATA[ABSTRACT Leishmaniasis is a disease caused by protozoan parasites of the genus Leishmania. These parasites infect to mammalian hosts, including canines that have been implicated as reservoirs of the parasite. The aim of this research was to standardize the technique of PCR-RFLP after amplification of the ITS1 region of Leishmania (Leishmania) infantum, as a useful tool for detection and molecular characterization. Promastigotes from culture and biopsies from dogs with visceral Leishmaniasis previously diagnosed by the Centro Antirrábico Nacional. The ITS1 region of the genomic DNA of Leishmania sp. was amplified using LITSR and L5,8S primers. The technique ITS1 PCR-RFLP applied, allowed the detection of Leishmania (L.) infantum in 10/10 of the isolates from parasites maintained in NNN culture medium, in 10/18 samples from spleen and 10/18 samples from popliteal lymph node. Optimal reaction conditions were 0.2 mM dNTPs, 0.1 pmol of each primer and 1U of Taq polymerase. The sensitivity of PCR was 3 ng/µL in isolates of parasites from NNN culture medium and 60 ng/µL in biopsy samples while the specificity was 100% for the detection of DNA of Leishmania sp. The restriction enzyme Hae III determined fragments of 184, 72 and 55 bp., which were specific to Leishmania (L.) infantum. The marker used is reliable for the detection and characterization of Leishmania sp. in dogs from endemic areas, which could be useful to verify the species of parasites circulating among animals.]]></p></abstract>
<kwd-group>
<kwd lng="es"><![CDATA[Leishmania infantum]]></kwd>
<kwd lng="es"><![CDATA[perros]]></kwd>
<kwd lng="es"><![CDATA[ITS -1 (Espaciador transcripto interno 1)]]></kwd>
<kwd lng="en"><![CDATA[Leishmania infantum]]></kwd>
<kwd lng="en"><![CDATA[dogs]]></kwd>
<kwd lng="en"><![CDATA[ITS1 (Internal Transcribed Spacer 1)]]></kwd>
</kwd-group>
</article-meta>
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