<?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>S0253-570X2018000200003</article-id>
<title-group>
<article-title xml:lang="es"><![CDATA[Desarrollo de un ensayo de PCR dúplex para la detección de Salmonella spp. y Staphylococcus aureus en leche cruda]]></article-title>
<article-title xml:lang="en"><![CDATA[Evaluation of a duplex PCR assay for the detection of Salmonella spp. and Staphylococcus aureus in raw milk]]></article-title>
</title-group>
<contrib-group>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Martínez-García]]></surname>
<given-names><![CDATA[Yuneilys Aleli]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Uffo-Reinoso]]></surname>
<given-names><![CDATA[Odalys]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Riverón-Alemán]]></surname>
<given-names><![CDATA[Yamilka]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Agüero-Fernández]]></surname>
<given-names><![CDATA[José Antonio]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Martínez-Vasallo]]></surname>
<given-names><![CDATA[Ailin]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
</contrib-group>
<aff id="Af1">
<institution><![CDATA[,Centro Nacional de Sanidad Agropecuaria  ]]></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>2018</year>
</pub-date>
<pub-date pub-type="epub">
<day>00</day>
<month>08</month>
<year>2018</year>
</pub-date>
<volume>40</volume>
<numero>2</numero>
<copyright-statement/>
<copyright-year/>
<self-uri xlink:href="http://scielo.sld.cu/scielo.php?script=sci_arttext&amp;pid=S0253-570X2018000200003&amp;lng=en&amp;nrm=iso"></self-uri><self-uri xlink:href="http://scielo.sld.cu/scielo.php?script=sci_abstract&amp;pid=S0253-570X2018000200003&amp;lng=en&amp;nrm=iso"></self-uri><self-uri xlink:href="http://scielo.sld.cu/scielo.php?script=sci_pdf&amp;pid=S0253-570X2018000200003&amp;lng=en&amp;nrm=iso"></self-uri><abstract abstract-type="short" xml:lang="es"><p><![CDATA[RESUMEN En alimentos contaminados, los patógenos de mayor importancia asociados a enfermedades de transmisión alimentaria (ETA), que producen brotes gastrointestinales agudos, se encuentran Salmonella spp. y Staphylococcus aureus. La causa fundamental de esta contaminación es la deficiente higiene en la obtención y manipulación de los mismos. En alimentos como la leche y derivados lácteos, las ETA alcanzan hasta 6 %. El presente trabajo tuvo como objetivos optimizar y estandarizar un ensayo de PCR dúplex para la detección de Salmonella spp. y Staphylococcus aureus en leche cruda. El método se basa en la amplificación simultánea del gen yfiR para Salmonella spp. y ARNr 23S para Staphylococcus aureus. Se determinaron las propiedades termodinámicas de cada cebador y se optimizaron los principales parámetros críticos del ensayo. El PCR dúplex optimizado demostró ser específico para los microorganismos diana y el ensayo detectó valores de hasta 1 pg/µL de ADN y 102ufc/mL en leche estéril. En el 46,7 % de las muestras de leche analizadas se detectó la presencia de S. aureus y no hubo muestra positiva a Salmonella spp. Los resultados sugieren que el PCR dúplex desarrollado es un ensayo sensible y específico para la detección simultánea de ambos patógenos en leche cruda.]]></p></abstract>
<abstract abstract-type="short" xml:lang="en"><p><![CDATA[ABSTRACT Infectious and toxigenic pathogens transmitted through food have been recognized as the cause of Foodborne Diseases (FBD). The main pathogens of concern are Salmonella spp. and Staphylococcus aureus. These bacteria are associated with acute gastrointestinal outbreaks caused by contaminated food. One of the causes of this contamination is the poor hygiene during the processing and handling of food. Foodborne outbreaks caused by milk and dairy products contaminated reach up to 6 %. The objective of the present work was to develop and standardize a duplex PCR assay for the detection of Salmonella spp. and Staphylococcus aureus in raw milk. The principle of the method is the simultaneous amplification of the genes yfiR for Salmonella spp. and ARNr 23S for Staphylococcus aureus. The thermodynamic properties of each primer were determined and the main critical parameters of the dPCR assay were optimized. The duplex PCR demonstrated its specificity for the target microorganisms and the detection limit of the assay was up to 1 pg/&#956;L of DNA and 102 cfu/mL in unpasteurized milk. In the milk samples analyzed, 46.7 % were contaminated with S. aureus and none of the samples were positive to Salmonella spp. The results suggest that the developed duplex PCR is sensible and specific for the simultaneous detection of both pathogens from raw milk.]]></p></abstract>
<kwd-group>
<kwd lng="es"><![CDATA[Salmonella spp.]]></kwd>
<kwd lng="es"><![CDATA[Staphylococcus aureus]]></kwd>
<kwd lng="es"><![CDATA[PCR dúplex]]></kwd>
<kwd lng="es"><![CDATA[leche cruda]]></kwd>
<kwd lng="en"><![CDATA[Salmonella spp.]]></kwd>
<kwd lng="en"><![CDATA[Staphylococcus aureus]]></kwd>
<kwd lng="en"><![CDATA[duplex PCR]]></kwd>
<kwd lng="en"><![CDATA[raw milk]]></kwd>
</kwd-group>
</article-meta>
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