<?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>S0253-570X2019000300002</article-id>
<title-group>
<article-title xml:lang="es"><![CDATA[Ensayo de PCR en tiempo real basado en SYBR-Green I para la detección de herpesvirus bovino tipo 1]]></article-title>
<article-title xml:lang="en"><![CDATA[SYBR Green I Based Real-Time RT-PCR assay for the detection of bovine herpesvirus type 1]]></article-title>
</title-group>
<contrib-group>
<contrib contrib-type="author">
<name>
<surname><![CDATA[García Delgado]]></surname>
<given-names><![CDATA[Laiyen]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Marrero-Perera]]></surname>
<given-names><![CDATA[Roxana]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Coronado Báez]]></surname>
<given-names><![CDATA[Liani]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Hinojosa López]]></surname>
<given-names><![CDATA[Yoandri]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Acevedo]]></surname>
<given-names><![CDATA[Ana María]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Relova Vento]]></surname>
<given-names><![CDATA[Damarys]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Vega Redondo]]></surname>
<given-names><![CDATA[Armando]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Perera González]]></surname>
<given-names><![CDATA[Carmen Laura]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
</contrib-group>
<aff id="Af1">
<institution><![CDATA[,Centro Nacional de Sanidad Agropecuaria Grupo de Virología Animal ]]></institution>
<addr-line><![CDATA[San José de las Lajas Mayabeque]]></addr-line>
<country>Cuba</country>
</aff>
<aff id="Af2">
<institution><![CDATA[,Centro Nacional de Sanidad Agropecuaria 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>12</month>
<year>2019</year>
</pub-date>
<pub-date pub-type="epub">
<day>00</day>
<month>12</month>
<year>2019</year>
</pub-date>
<volume>41</volume>
<numero>3</numero>
<copyright-statement/>
<copyright-year/>
<self-uri xlink:href="http://scielo.sld.cu/scielo.php?script=sci_arttext&amp;pid=S0253-570X2019000300002&amp;lng=en&amp;nrm=iso"></self-uri><self-uri xlink:href="http://scielo.sld.cu/scielo.php?script=sci_abstract&amp;pid=S0253-570X2019000300002&amp;lng=en&amp;nrm=iso"></self-uri><self-uri xlink:href="http://scielo.sld.cu/scielo.php?script=sci_pdf&amp;pid=S0253-570X2019000300002&amp;lng=en&amp;nrm=iso"></self-uri><abstract abstract-type="short" xml:lang="es"><p><![CDATA[RESUMEN El herpesvirus bovino tipo 1 (BoHV-1) infecta los tractos respiratorio y genital del ganado bovino y varias especies de rumiantes. La infección puede resultar en diferentes enfermedades como rinotraqueitis infecciosa bovina, vulvovaginitis pustular infecciosa en hembras y balanopostitis pustular infecciosa en machos, con un severo impacto económico en la producción ganadera. La infección produce diferentes cuadros clínicos como respiratorios, conjuntivitis, abortos, enteritis e infección sistémica fatal en terneros recién nacidos. Una de las características más importante del herpesvirus bovino tipo 1 es el establecimiento de una infección latente que puede ser reactivada bajo condiciones de estrés o por tratamientos con altas dosis de corticosteroides. El presente trabajo tuvo como objetivo el desarrollo de un ensayo de PCR en tiempo real basado en SYBR-Green I para la detección de herpesvirus bovino 1, mediante la amplificación de un fragmento del gen de la timidina quinasa viral. Se establecieron los parámetros de los componentes críticos de la reacción de PCR, así como de las condiciones termodinámicas. Mediante el análisis de la curva de disociación se verificó que las curvas de amplificación fueron específicas (Tm = 93°C ± 1°C) y que no hubo amplificaciones inespecíficas. Se obtuvieron altos niveles de especificidad, repetibilidad y sensibilidad, con un límite de detección de 100.5 DICT50/mL. Se evaluó la sensibilidad diagnóstica a partir de exudados nasales procedentes de animales de una reproducción experimental, de reactivación de la infección latente y de vaquerías experimentales. El ensayo de PCR en tiempo real con SYBR-Green I demostró ser más sensible para el diagnóstico de BoHV-1, que el ensayo de PCR en punto final y el aislamiento viral.]]></p></abstract>
<abstract abstract-type="short" xml:lang="en"><p><![CDATA[ABSTRACT Bovine herpesvirus type 1 (BoHV-1) infects the respiratory and genital tracts of cattle and several species of ruminants. The infection can result in different diseases such as infectious bovine rhinotracheitis, infectious pustular vulvovaginitis in females and infectious pustular balanoposthitis in males, with a severe economic impact on livestock production. The infection causes respiratory symptoms, conjunctivitis, abortions, enteritis, and fatal systemic infection in newborn calves. One of the most important characteristics of bovine herpesvirus type 1 is the establishment of a latent infection that can be reactivated under stress conditions or by treatment with high doses of corticosteroids. The present work was aimed at developing an SYBR Green I Based Real-Time RT-PCR assay for the detection of bovine herpesvirus type 1, by amplifying a fragment of the viral thymidine kinase gene. The parameters of the critical components of the PCR reaction as well as the thermodynamic conditions were established. Dissociation curve analysis showed that the amplification curves were specific (Tm = 93°C ± 1°C) and that there were no unspecific amplifications. High levels of specificity, repeatability and sensitivity were obtained, with a detection limit of 100.5 TCID50/mL. Diagnostic sensitivity was assessed on the basis of nasal exudates from experimental breeding animals, reactivation of latent infection and experimental dairy farms. The SYBR Green I Based Real-Time RT-PCR assay proved to be more sensitive for the diagnosis of BoHV-1 than the endpoint PCR assay and virus isolation.]]></p></abstract>
<kwd-group>
<kwd lng="es"><![CDATA[herpesvirus bovino tipo 1]]></kwd>
<kwd lng="es"><![CDATA[PCR en tiempo real]]></kwd>
<kwd lng="es"><![CDATA[SYBR-Green I]]></kwd>
<kwd lng="en"><![CDATA[bovine herpesvirus type 1]]></kwd>
<kwd lng="en"><![CDATA[real-time PCR]]></kwd>
<kwd lng="en"><![CDATA[SYBR-Green I]]></kwd>
</kwd-group>
</article-meta>
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