<?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>0253-570X</journal-id>
<journal-title><![CDATA[Revista de Salud Animal]]></journal-title>
<abbrev-journal-title><![CDATA[Rev Salud Anim.]]></abbrev-journal-title>
<issn>0253-570X</issn>
<publisher>
<publisher-name><![CDATA[Centro Nacional de Sanidad Agropecuaria]]></publisher-name>
</publisher>
</journal-meta>
<article-meta>
<article-id>S0253-570X2020000200003</article-id>
<title-group>
<article-title xml:lang="es"><![CDATA[Desarrollo de PCR especie-específicos para la detección de micoplasmas contaminantes de cultivos celulares]]></article-title>
<article-title xml:lang="en"><![CDATA[Development of species-specific PCRs for the detection of mycoplasmas cell culture contaminants]]></article-title>
</title-group>
<contrib-group>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Pérez Castillo]]></surname>
<given-names><![CDATA[Anisleidy]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Duque Ortiz]]></surname>
<given-names><![CDATA[Arianna]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Burgher Pulgarón]]></surname>
<given-names><![CDATA[Yaima]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Agüero Fernández]]></surname>
<given-names><![CDATA[Jose Antonio]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Solis Castells]]></surname>
<given-names><![CDATA[Mario Sergio]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Lobo-Rivero]]></surname>
<given-names><![CDATA[Evelyn]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
</contrib-group>
<aff id="Af1">
<institution><![CDATA[,Centro Nacional de Sanidad Agropecuaria Dirección de Salud Animal Grupo de Bacteriología-Parasitología]]></institution>
<addr-line><![CDATA[San José de las Lajas Mayabeque]]></addr-line>
<country>Cuba</country>
</aff>
<pub-date pub-type="pub">
<day>00</day>
<month>08</month>
<year>2020</year>
</pub-date>
<pub-date pub-type="epub">
<day>00</day>
<month>08</month>
<year>2020</year>
</pub-date>
<volume>42</volume>
<numero>2</numero>
<copyright-statement/>
<copyright-year/>
<self-uri xlink:href="http://scielo.sld.cu/scielo.php?script=sci_arttext&amp;pid=S0253-570X2020000200003&amp;lng=en&amp;nrm=iso"></self-uri><self-uri xlink:href="http://scielo.sld.cu/scielo.php?script=sci_abstract&amp;pid=S0253-570X2020000200003&amp;lng=en&amp;nrm=iso"></self-uri><self-uri xlink:href="http://scielo.sld.cu/scielo.php?script=sci_pdf&amp;pid=S0253-570X2020000200003&amp;lng=en&amp;nrm=iso"></self-uri><abstract abstract-type="short" xml:lang="es"><p><![CDATA[RESUMEN La contaminación por Mollicutes en los cultivos celulares es frecuente. Especies como Mycoplasma arginini, Mycoplasma salivarium, Mycoplasma fermentans, Mycoplasma hyorhinis, Mycoplasma orale y Acholeplasma laidlawii se identifican como causantes del 95 % de las contaminaciones. El presente trabajo tuvo como objetivo desarrollar PCR especie-específicos para la identificación de las especies de micoplasmas contaminantes más frecuentes en cultivos celulares. Se optimizaron los parámetros críticos de la PCR y se determinaron la sensibilidad y especificidad analítica. Se estableció como valor óptimo de temperatura de hibridación 55ºC; concentración de magnesio 1,5 mM y de cebadores se seleccionó un rango 0,3-0,5 µM. La sensibilidad analítica de la PCR fue de 1 pg/µL, mientras que su especificidad analítica fue 100 %. Se realizó la PCR especie-específico a 58 muestras de cultivo celulares donde previamente se detectó la presencia de Mollicutes. Como resultado se detectó Mycoplasma orale como la especie más frecuente, seguida de Mycoplasma fermentans, Acholeplasma laidlawii, Mycoplasma arginini, Mycoplasma hyorhinis y Mycoplasma salivarium. Los resultados de este estudio confirman que los ensayos PCR especie-específicos desarrollados son factibles de emplear para la identificación de especies de micoplasmas en cultivos celulares.]]></p></abstract>
<abstract abstract-type="short" xml:lang="en"><p><![CDATA[ABSTRACT Mollicute contamination in cell cultures is common. Species such as Mycoplasma arginini, Mycoplasma salivarium, Mycoplasma fermentans, Mycoplasma hyorhinis, Mycoplasma orale, and Acholeplasma laidlawii are identified as causing 95 % of the contaminations. The present work is aimed at developing species-specific PCRs for the identification of the most frequent contaminating mycoplasma species in cell cultures. The critical parameters of PCR were optimized and the analytical sensitivity and specificity were determined. The optimum temperature for hybridization was set at 55ºC; magnesium concentration at 1,5 mM. A range of 0,3-0,5 µM was selected for the primers. The analytical sensitivity of PCR was 1 pg/µL, while its analytical specificity was 100 %. Species-specific PCR was performed on 58 cell culture samples where the presence of Mollicutes was previously detected. As a result, Mycoplasma orale was detected as the most frequent species, followed by Mycoplasma fermentans, Acholeplasma laidlawii, Mycoplasma arginini, Mycoplasma hyorhinis, and Mycoplasma salivarium. The results of this study confirm that the species-specific PCR assays developed are feasible to use for the identification of mycoplasma species in cell cultures.]]></p></abstract>
<kwd-group>
<kwd lng="es"><![CDATA[cultivos celulares]]></kwd>
<kwd lng="es"><![CDATA[diagnóstico]]></kwd>
<kwd lng="es"><![CDATA[micoplasmas]]></kwd>
<kwd lng="es"><![CDATA[PCR]]></kwd>
<kwd lng="en"><![CDATA[cell culture]]></kwd>
<kwd lng="en"><![CDATA[diagnosis]]></kwd>
<kwd lng="en"><![CDATA[mycoplasmas]]></kwd>
<kwd lng="en"><![CDATA[PCR]]></kwd>
</kwd-group>
</article-meta>
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