<?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>1010-2752</journal-id>
<journal-title><![CDATA[Revista de Protección Vegetal]]></journal-title>
<abbrev-journal-title><![CDATA[Rev. Protección Veg.]]></abbrev-journal-title>
<issn>1010-2752</issn>
<publisher>
<publisher-name><![CDATA[Centro Nacional de Sanidad Agropecuaria]]></publisher-name>
</publisher>
</journal-meta>
<article-meta>
<article-id>S1010-27522019000300005</article-id>
<title-group>
<article-title xml:lang="en"><![CDATA[Evaluation of a non-radioactive dot-blot nucleic acid hybridization as a detection tool for citrus leaf blotch virus in Cuba]]></article-title>
<article-title xml:lang="es"><![CDATA[Evaluación de la hibridación de ácidos nucleicos no radiactivo dot-blot como herramienta de detección para citrus leaf blotch virus en Cuba]]></article-title>
</title-group>
<contrib-group>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Hernández-Rodríguez]]></surname>
<given-names><![CDATA[Lester]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
<xref ref-type="aff" rid="Aaf"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[García-García]]></surname>
<given-names><![CDATA[Gabriel]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
<xref ref-type="aff" rid="Aaf"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Pérez-Castro]]></surname>
<given-names><![CDATA[Juana María]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Batista-Le Riverend]]></surname>
<given-names><![CDATA[Lochy]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Zamora-Rodríguez]]></surname>
<given-names><![CDATA[Victoria]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Peña-Barzaga]]></surname>
<given-names><![CDATA[Inés]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Ramos-González]]></surname>
<given-names><![CDATA[Pedro Luis]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
<xref ref-type="aff" rid="Aaf"/>
</contrib>
</contrib-group>
<aff id="Af1">
<institution><![CDATA[,Instituto de Investigaciones en Fruticultura Tropical  ]]></institution>
<addr-line><![CDATA[Playa La Habana]]></addr-line>
<country>Cuba</country>
</aff>
<aff id="Af2">
<institution><![CDATA[,Instituto Nacional de Investigación Agropecuaria Estación Experimental INIA Salto Grande ]]></institution>
<addr-line><![CDATA[Salto ]]></addr-line>
<country>Uruguay</country>
</aff>
<aff id="Af3">
<institution><![CDATA[,Analytical Food Laboratories  ]]></institution>
<addr-line><![CDATA[ ]]></addr-line>
</aff>
<aff id="Af4">
<institution><![CDATA[,Instituto Biológico de São Paulo  ]]></institution>
<addr-line><![CDATA[São Paulo ]]></addr-line>
<country>Brazil</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>34</volume>
<numero>3</numero>
<copyright-statement/>
<copyright-year/>
<self-uri xlink:href="http://scielo.sld.cu/scielo.php?script=sci_arttext&amp;pid=S1010-27522019000300005&amp;lng=en&amp;nrm=iso"></self-uri><self-uri xlink:href="http://scielo.sld.cu/scielo.php?script=sci_abstract&amp;pid=S1010-27522019000300005&amp;lng=en&amp;nrm=iso"></self-uri><self-uri xlink:href="http://scielo.sld.cu/scielo.php?script=sci_pdf&amp;pid=S1010-27522019000300005&amp;lng=en&amp;nrm=iso"></self-uri><abstract abstract-type="short" xml:lang="en"><p><![CDATA[ABSTRACT A non-radioactive dot-blot nucleic acid hybridization method was evaluated for detecting citrus leaf blotch virus (CLBV, Flexiviridae: Citrivirus) in total RNA extracts from kumquat trees. DNA fragments partially encompassing the open reading frames encoding the polymerase (RdRp) and the movement (MP coat (CP) proteins of CLBV were used to generate digoxigenin (DIG-11-dUTP) probes by PCR. Probes were individually generated using cloned DNA fragments of the Cuban CLBV isolate as templates. In the assays, several working conditions, including the use of different membranes, hybridization and washing buffers were tested. The method detected up to 40 pg of a plasmid containing the complete CLBV genome, as well as viral genomic sequences in RNA extracts of naturally infected trees of 'Nagami' kumquat (Fortunella margarita) and 'Round' kumquat (F. japonica). Results support considering the method developed an appropriated tool to be used after its validation by the programs of sanitation, quarantine, and certification of citrus propagation materials in Cuba.]]></p></abstract>
<abstract abstract-type="short" xml:lang="es"><p><![CDATA[RESUMEN Se evaluó un método de hibridación de ácidos nucleicos no radioactivo en formato dot-blot para la detección precisa de citrus leaf blotch virus (CLBV: Citrivirus) en extractos de ARN totales de árboles de cítricos. Los fragmentos de ADN que conforman parcialmente los marcos de lectura abiertos que codifican la polimerasa (RdRp), la proteína de movimiento (MP) y la proteína de la cubierta (CP) de CLBV se utilizaron para generar sondas marcadas con digoxigenina (DIG-11-dUTP) mediante PCR. Para generar las sondas se utilizaron fragmentos de ADN clonados a partir del aislado de CLBV de Cuba. Se evaluaron varias condiciones de trabajo que incluyeron el uso de diferentes membranas, soluciones de hibridación y tampones de lavado. Con el método evaluado fue posible detectar hasta 40 pg de un plasmidio que contiene el genoma completo de CLBV, así como material genómico del virus en extractos de ARN de plantas de kumquat 'Nagami' (Fortunella margarita) y kumquat 'Round' (F. japonica) infectadas de forma natural con el virus. Los resultados evidenciaron que el método desarrollado es una herramienta útil y altamente recomendable, una vez sea validado, para su uso en los programas de saneamiento, cuarentena y certificación del material de propagación de cítricos en Cuba.]]></p></abstract>
<kwd-group>
<kwd lng="en"><![CDATA[budwood]]></kwd>
<kwd lng="en"><![CDATA[certification]]></kwd>
<kwd lng="en"><![CDATA[Citrivirus]]></kwd>
<kwd lng="en"><![CDATA[CLBV]]></kwd>
<kwd lng="en"><![CDATA[diagnosis]]></kwd>
<kwd lng="en"><![CDATA[Flexiviridae]]></kwd>
<kwd lng="en"><![CDATA[Fortunella]]></kwd>
<kwd lng="es"><![CDATA[certificación]]></kwd>
<kwd lng="es"><![CDATA[CLBV]]></kwd>
<kwd lng="es"><![CDATA[Citrivirus]]></kwd>
<kwd lng="es"><![CDATA[Flexiviridae]]></kwd>
<kwd lng="es"><![CDATA[diagnóstico]]></kwd>
<kwd lng="es"><![CDATA[Fortunella]]></kwd>
<kwd lng="es"><![CDATA[material de propagación]]></kwd>
</kwd-group>
</article-meta>
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