<?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>S0253-570X2018000300001</article-id>
<title-group>
<article-title xml:lang="en"><![CDATA[Standarization of a SYBR Green-I based real time RT-PCR assay for the detection of equine influenza virus]]></article-title>
<article-title xml:lang="es"><![CDATA[Estandarización de un ensayo de RT-PCR en tiempo real basado en SYBR Green-I para la detección del virus de la influenza equina]]></article-title>
</title-group>
<contrib-group>
<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[Lazo]]></surname>
<given-names><![CDATA[Ana María]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Relova]]></surname>
<given-names><![CDATA[Damarys]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Perera]]></surname>
<given-names><![CDATA[Carmen Laura]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
</contrib-group>
<aff id="Af1">
<institution><![CDATA[,National Center for Animal and Plant Health Animal Virology Group ]]></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>2018</year>
</pub-date>
<pub-date pub-type="epub">
<day>00</day>
<month>12</month>
<year>2018</year>
</pub-date>
<volume>40</volume>
<numero>3</numero>
<copyright-statement/>
<copyright-year/>
<self-uri xlink:href="http://scielo.sld.cu/scielo.php?script=sci_arttext&amp;pid=S0253-570X2018000300001&amp;lng=en&amp;nrm=iso"></self-uri><self-uri xlink:href="http://scielo.sld.cu/scielo.php?script=sci_abstract&amp;pid=S0253-570X2018000300001&amp;lng=en&amp;nrm=iso"></self-uri><self-uri xlink:href="http://scielo.sld.cu/scielo.php?script=sci_pdf&amp;pid=S0253-570X2018000300001&amp;lng=en&amp;nrm=iso"></self-uri><abstract abstract-type="short" xml:lang="en"><p><![CDATA[ABSTRACT Equine influenza is an acute respiratory infection of horses, donkeys, mules and zebras. Nowadays, the reverse transcriptase polymerase chain reaction (RT-PCR) and real-time RT-PCR (rRT-PCR) assays are being widely used in diagnostic laboratories as a more sensitive alternative to virus isolation for the detection of equine influenza virus. The purpose of this study was to standardize an rRT-PCR, based on SYBR Green-I coupled with melting curve analysis for the detection of equine influenza virus (gene M) and the typing of hemagglutinin gene (HA). Sensitivity and specificity of the assay (gene M) were evaluated. In terms of the matrix gene copy number, the detection limit was 4.5 gene copies/&#956;l of in vitro transcribed RNA. The linear range obtained for the transcript in the assay genered a typical standard curve from 107 until 100 gene copies/µl in terms of RNA copy number. The correlation coefficient (R2) was 0.99 for the RNA copies detected in nuclease free water and the error of standard curve was 0.0139. The rRT-PCR assay showed to be specific for equine influenza virus. No specific amplification curves were obtained with any of the other viruses tested such as equine viral arteritis, equine herpesvirus-1 and equine herpesvirus-4. The assay showed amplification and specific melting curves when the second set of primers against HA gene was evaluated. Thus, the proposed SYBR Green-I based rRT-PCR assay for the detection of equine influenza virus can be used in case of a possible emergency of this agent in Cuba.]]></p></abstract>
<abstract abstract-type="short" xml:lang="es"><p><![CDATA[RESUMEN La influenza equina es una infección respiratoria aguda de los caballos, burros, mulas y cebras. Actualmente, los ensayos de reverso transcripción acoplados a la reacción en cadena de la polimerasa (RT-PCR) y RT-PCR en tiempo real (rRT-PCR) están siendo ampliamente usados en los laboratorios de diagnóstico, como una alternativa más sensible al aislamiento viral, para la detección del virus de influenza equina. El objetivo de este estudio fue estandarizar un ensayo de rRT-PCR, basado en SYBR Green-I acoplado a análisis de curvas de disociación para la detección del virus de influenza equina (gen M) y la tipificación del gen de la hemaglutinina (HA). Se evaluaron la sensibilidad y la especificidad del ensayo (gen M). En términos de números de copias del gen de la matriz, el límite de detección fue de 4,5 copias del gen/&#956;L del ARN transcripto in vitro. El rango lineal obtenido para el transcripto en el ensayo generó una curva estándar desde 107 hasta 100 copias del gen/µL en término de número de copias de ARN. El coeficiente de correlación (R2) fue de 0,99 para las copias de ARN detectadas en agua libre de nucleasas. El error de la curva estándar fue de 0,0139. El ensayo de rRT-PCR mostró ser específico para el virus de influenza equina. No se obtuvieron curvas de amplificación específicas con los otros virus evaluados, como son arteritis viral equina, herpesvirus equino-1 y herpesvirus equino-4. El ensayo mostró amplificación y curvas de disociación específicas cuando se evaluó la segunda pareja de cebadores contra el gen de la hemaglutinina. Así, el ensayo propuesto de rRT-PCR, basado en SYBR Green-I para la detección del virus de influenza equina, puede ser usado en caso de una posible emergencia de este agente en Cuba.]]></p></abstract>
<kwd-group>
<kwd lng="en"><![CDATA[equine influenza virus]]></kwd>
<kwd lng="en"><![CDATA[SYBR Green-I based real time RT-PCR assay]]></kwd>
<kwd lng="es"><![CDATA[virus de la influenza equina]]></kwd>
<kwd lng="es"><![CDATA[ensayo de RT-PCR en tiempo real basado en SYBR Green-I]]></kwd>
</kwd-group>
</article-meta>
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