<?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>0375-0760</journal-id>
<journal-title><![CDATA[Revista Cubana de Medicina Tropical]]></journal-title>
<abbrev-journal-title><![CDATA[Rev Cubana Med Trop]]></abbrev-journal-title>
<issn>0375-0760</issn>
<publisher>
<publisher-name><![CDATA[Centro Nacional de Información de Ciencias Médicas]]></publisher-name>
</publisher>
</journal-meta>
<article-meta>
<article-id>S0375-07602021000300007</article-id>
<title-group>
<article-title xml:lang="es"><![CDATA[Identificación de especies de Leishmania mediante PCR en tiempo real acoplada a curvas de fusión de alta resolución]]></article-title>
<article-title xml:lang="en"><![CDATA[Identification of Leishmania species by real time PCR coupled with high resolution melting curve analysis]]></article-title>
</title-group>
<contrib-group>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Villarreal Julio]]></surname>
<given-names><![CDATA[Rafael Guillermo]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Herrera]]></surname>
<given-names><![CDATA[Giovanny]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Muskus López]]></surname>
<given-names><![CDATA[Carlos E.]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
</contrib-group>
<aff id="Af1">
<institution><![CDATA[,Universidad de Antioquia Facultad de Medicina ]]></institution>
<addr-line><![CDATA[ ]]></addr-line>
<country>Colombia</country>
</aff>
<pub-date pub-type="pub">
<day>00</day>
<month>12</month>
<year>2021</year>
</pub-date>
<pub-date pub-type="epub">
<day>00</day>
<month>12</month>
<year>2021</year>
</pub-date>
<volume>73</volume>
<numero>3</numero>
<copyright-statement/>
<copyright-year/>
<self-uri xlink:href="http://scielo.sld.cu/scielo.php?script=sci_arttext&amp;pid=S0375-07602021000300007&amp;lng=en&amp;nrm=iso"></self-uri><self-uri xlink:href="http://scielo.sld.cu/scielo.php?script=sci_abstract&amp;pid=S0375-07602021000300007&amp;lng=en&amp;nrm=iso"></self-uri><self-uri xlink:href="http://scielo.sld.cu/scielo.php?script=sci_pdf&amp;pid=S0375-07602021000300007&amp;lng=en&amp;nrm=iso"></self-uri><abstract abstract-type="short" xml:lang="es"><p><![CDATA[RESUMEN  Introducción:  La leishmaniasis es una enfermedad causada por parásitos del género Leishmania. En Colombia se han informado 10 especies patógenas. El diagnóstico parasitológico tradicional basado en la observación de los parásitos no permite identificar la especie, por lo cual se deben emplear métodos moleculares, entre ellos la reacción en cadena de la polimerasa o PCR convencional, pero esta presenta algunas limitaciones y requiere extensos periodos de tiempo para la obtención de resultados, que en ocasiones no son concluyentes.  Objetivo:  Evaluar un método basado en PCR en tiempo real acoplado a curva de temperatura de desnaturalización media de alta resolución (PCR-HRM) que permita el diagnóstico y la identificación simultánea de parásitos del género Leishmania en muestras clínicas de humanos y en cultivos in vitro de manera sensible y específica.  Métodos:  Se estandarizó una PCR-HRM, mediante la cual se evaluaron 237 muestras clínicas, 98 clasificadas como positivas y 139 como negativas, parasitológicamente por directo y/o cultivo. Las tipificaciones fueron comparadas con los resultados en paralelo obtenidos de una variante de la PCR, realizando cortes al amplicon que generó un fragmento de restricción de longitud polimórfica o PCR-RFLP que había sido previamente estandarizada.  Resultados:  Se logró implementar una PCR-HRM para el diagnóstico e identificación de especies de Leishmania, logrando un 100 % de concordancia con las tipificaciones obtenidas por PCR-RFLP. Incluso, se logró detectar e identificar el parásito en muestras diagnosticadas como negativas por los métodos convencionales. Se encontró que con un porcentaje de confiabilidad superior al 95 %, se lograron tipificar 91 muestras de 98; de estas el 81,63 % de los casos fueron L. panamensis, el 11,22% L. braziliensis e indeterminadas el 7,14 % de los casos.  Conclusiones:  La PCR-HRM es un buen método que permite la identificación de las especies más prevalentes en Colombia, comparando temperaturas medias de desnaturalización específicas según la especie de Leishmania involucrada.]]></p></abstract>
<abstract abstract-type="short" xml:lang="en"><p><![CDATA[ABSTRACT  Introduction:  Leishmaniasis is a disease caused by parasites of the genus Leishmania. Ten pathogenic species have been reported in Colombia. Traditional parasite diagnosis based on observation of the parasites does not make it possible to identify the species. Therefore, it is necessary to use molecular methods, among them conventional polymerase chain reaction or PCR, but this test presents some limitations and requires long periods of time to obtain results which sometimes are not conclusive.  Objective:  Evaluate a method based on real time PCR coupled with high resolution mean denaturalization temperature curve analysis (HRM-PCR) for the diagnosis and simultaneous identification of parasites of the genus Leishmania in clinical samples from humans and in vitro cultures in a sensitive and specific manner.  Methods:  Standardization was performed of an HRM-PCR with which 237 clinical samples were evaluated, 98 classified as positive and 139 as negative, by direct parasitological examination and/or culture. The typing obtained was compared with parallel results from a PCR variant, making cuts on the amplicon that generated a restriction fragment length polymorphism or PCR-RFLP previously standardized.  Results:  An HRM-PCR could be implemented for the diagnosis and identification of Leishmania species, achieving 100% concordance with the typing obtained by PCR-RFLP. It was even possible to detect and identify the parasite in samples diagnosed as negative by conventional methods. Of the total 98 samples, 91 could be typed with a percentage of reliability above 95%. Of these, 81.63% of the cases were L. panamensis, 11.22% were L. braziliensis and 7.14 % were indeterminate.  Conclusions:  HRM-PCR is a good method to identify the species most prevalent in Colombia, comparing specific mean denaturalization temperatures according to the Leishmania species involved.]]></p></abstract>
<kwd-group>
<kwd lng="es"><![CDATA[Leishmania]]></kwd>
<kwd lng="es"><![CDATA[diagnóstico]]></kwd>
<kwd lng="es"><![CDATA[identificación]]></kwd>
<kwd lng="es"><![CDATA[PCR en tiempo real]]></kwd>
<kwd lng="es"><![CDATA[HRM]]></kwd>
<kwd lng="en"><![CDATA[Leishmania]]></kwd>
<kwd lng="en"><![CDATA[diagnosis]]></kwd>
<kwd lng="en"><![CDATA[identification]]></kwd>
<kwd lng="en"><![CDATA[real time PCR]]></kwd>
<kwd lng="en"><![CDATA[HRM]]></kwd>
</kwd-group>
</article-meta>
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