<?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>S0253-570X2023000100005</article-id>
<title-group>
<article-title xml:lang="es"><![CDATA[Ensayo de PCR en tiempo real basado en SYBR Green para la detección del virus de la diarrea epidémica porcina]]></article-title>
<article-title xml:lang="en"><![CDATA[SYBR Green-based real-time PCR assay for the detection of porcine epidemic diarrhea virus]]></article-title>
</title-group>
<contrib-group>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Granda Moncayo]]></surname>
<given-names><![CDATA[Danilo]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Proaño Pérez]]></surname>
<given-names><![CDATA[Freddy]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Garrido Haro]]></surname>
<given-names><![CDATA[Ana D.]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Barrera Valle]]></surname>
<given-names><![CDATA[Maritza]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
</contrib-group>
<aff id="Af1">
<institution><![CDATA[,Escuela Superior Politécnica del Ejército Departamento Ciencias de la Vida ]]></institution>
<addr-line><![CDATA[ ]]></addr-line>
<country>Ecuador</country>
</aff>
<aff id="Af2">
<institution><![CDATA[,Agencia de Regulación y Control Fito y Zoosanitario Laboratorio de Biología Molecular ]]></institution>
<addr-line><![CDATA[ Quito]]></addr-line>
<country>Ecuador</country>
</aff>
<aff id="Af3">
<institution><![CDATA[,Universidad Técnica de Manabí Facultad de Ciencias Veterinarias ]]></institution>
<addr-line><![CDATA[ Portoviejo]]></addr-line>
<country>Ecuador</country>
</aff>
<pub-date pub-type="pub">
<day>00</day>
<month>00</month>
<year>2023</year>
</pub-date>
<pub-date pub-type="epub">
<day>00</day>
<month>00</month>
<year>2023</year>
</pub-date>
<volume>45</volume>
<copyright-statement/>
<copyright-year/>
<self-uri xlink:href="http://scielo.sld.cu/scielo.php?script=sci_arttext&amp;pid=S0253-570X2023000100005&amp;lng=en&amp;nrm=iso"></self-uri><self-uri xlink:href="http://scielo.sld.cu/scielo.php?script=sci_abstract&amp;pid=S0253-570X2023000100005&amp;lng=en&amp;nrm=iso"></self-uri><self-uri xlink:href="http://scielo.sld.cu/scielo.php?script=sci_pdf&amp;pid=S0253-570X2023000100005&amp;lng=en&amp;nrm=iso"></self-uri><abstract abstract-type="short" xml:lang="es"><p><![CDATA[RESUMEN La diarrea epidémica porcina (DEP) es una enfermedad contagiosa producida por el virus del mismo nombre que es clasificado dentro del género Alphacoronavirus, familia Coronaviridae. Este virus produce diarrea, vómitos y pérdida de peso en cerdos de todas las edades, pero en los cerditos neonatos alcanza una morbilidad y mortalidad de hasta el 100%. En Ecuador, fue diagnosticada por primera vez en el 2014 mediante PCR convencional o en punto final. Actualmente existe la necesidad de contar con un ensayo ultrasensible, específico, que permita trabajar un gran número de muestras para facilitar la vigilancia epidemiológica de nuevos brotes y detectar animales portadores. Se desarrolló y evaluó un ensayo de PCR en tiempo real con un colorante intercalador de ADN como el SYBR Green para detectar el virus de la DEP (VDEP) en heces de cerdos, con cebadores que tienen como diana el gen N del virus. En la evaluación de la sensibilidad analítica se utilizó un patrón de ARN del VDEP sintetizado en el laboratorio, determinándose que el límite de detección del ensayo desarrollado fue de 4 copias de ARN del VDEP, comparando con la PCR en punto final que fue 100 veces más sensible, la especificidad del ensayo se basó en la posibilidad de detectar falsos positivos mediante la curva de disociación. Los valores de Ct (valor umbral) en el análisis intra e inter-ensayo mostraron coeficientes de variación válidos. En la evaluación del desempeño diagnóstico, el ensayo detectó 18 muestras positivas de un total de 52 para un 100% de concordancia con el PCR en punto final. Se concluye que el ensayo de PCR en tiempo real con SYBR Green desarrollado es altamente sensible, específico, reproducible y rápido para el diagnóstico del VDEP en homogenados de mucosa intestinal de cerdos.]]></p></abstract>
<abstract abstract-type="short" xml:lang="en"><p><![CDATA[ABSTRACT Porcine epidemic diarrhea (PED) is a contagious disease caused by its virus, classified within the genus Alphacoronavirus, family Coronaviridae. This virus causes diarrhea, vomiting and weight loss in pigs of all ages, especially in neonatal piglets, with morbidity and mortality of up to 100%. In Ecuador, the disease was diagnosed for the first time in 2014 by conventional or endpoint PCR. Currently, there is a need for an ultrasensitive and specific assay, which allows working a large number of samples to facilitate epidemiological surveillance of new outbreaks and detect carrier animals. A real-time PCR assay intercalating DNA dyes such as SYBR Green was developed and assessed to detect PED virus (PEDv) in swine feces with primers targeting the N-gene of the virus. A PEDv RNA standard synthesized in the laboratory was used in the assessment of the analytical sensitivity. The detection limit of the developed assay was determined to be 4 copies of PEDv RNA, compared to endpoint PCR which was 100 times more sensitive. Assay specificity was based on the possibility of detecting false positives using the melting curve. Cycle threshold (Ct) values in intra- and inter-assay analysis showed valid coefficients of variation. When assessing diagnostic performance, the assay detected 18 positive samples out of a total of 52 for 100 % agreement with the endpoint PCR. It is concluded that the developed SYBR Green-based real-time PCR assay is highly sensitive, specific, reproducible and rapid for the detection of PEDv in swine intestinal mucosa homogenates.]]></p></abstract>
<kwd-group>
<kwd lng="es"><![CDATA[PCR en tiempo real]]></kwd>
<kwd lng="es"><![CDATA[SYBR Green]]></kwd>
<kwd lng="es"><![CDATA[diarrea epidémica porcina]]></kwd>
<kwd lng="es"><![CDATA[gen N]]></kwd>
<kwd lng="en"><![CDATA[real-time PCR]]></kwd>
<kwd lng="en"><![CDATA[SYBR Green]]></kwd>
<kwd lng="en"><![CDATA[porcine epidemic diarrhea]]></kwd>
<kwd lng="en"><![CDATA[N gene]]></kwd>
</kwd-group>
</article-meta>
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