<?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>0120-0690</journal-id>
<journal-title><![CDATA[Revista Colombiana de Ciencias Pecuarias]]></journal-title>
<abbrev-journal-title><![CDATA[Rev Colom Cienc Pecua]]></abbrev-journal-title>
<issn>0120-0690</issn>
<publisher>
<publisher-name><![CDATA[Facultad de Ciencias Agrarias, Universidad de Antioquia]]></publisher-name>
</publisher>
</journal-meta>
<article-meta>
<article-id>S0120-06902018000400285</article-id>
<article-id pub-id-type="doi">10.17533/udea.rccp.v31n4a05</article-id>
<title-group>
<article-title xml:lang="en"><![CDATA[Design and analytical validation of a duplex PCR for Ehrlichia and Rickettsia detection in ticks]]></article-title>
<article-title xml:lang="es"><![CDATA[Diseño y validación analítica de una PCR dúplex para la detección de Ehrlichia y Rickettsia en garrapatas.]]></article-title>
<article-title xml:lang="pt"><![CDATA[Projeto e validação analítica de uma PCR duplex para detecção de Ehrlichia e Rickettsia em carrapatos]]></article-title>
</title-group>
<contrib-group>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Pérez Pérez]]></surname>
<given-names><![CDATA[Juan C]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Montoya Ruiz]]></surname>
<given-names><![CDATA[Carolina]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Arroyave Sierra]]></surname>
<given-names><![CDATA[Esteban]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Paternina]]></surname>
<given-names><![CDATA[Luis E]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Rodas]]></surname>
<given-names><![CDATA[Juan D]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
</contrib-group>
<aff id="Af1">
<institution><![CDATA[,Universidad de Antioquia  ]]></institution>
<addr-line><![CDATA[Medellín ]]></addr-line>
<country>Colombia</country>
</aff>
<aff id="Af2">
<institution><![CDATA[,Universidad de Sucre  ]]></institution>
<addr-line><![CDATA[Sincelejo ]]></addr-line>
<country>Colombia</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>31</volume>
<numero>4</numero>
<fpage>285</fpage>
<lpage>294</lpage>
<copyright-statement/>
<copyright-year/>
<self-uri xlink:href="http://www.scielo.org.co/scielo.php?script=sci_arttext&amp;pid=S0120-06902018000400285&amp;lng=en&amp;nrm=iso"></self-uri><self-uri xlink:href="http://www.scielo.org.co/scielo.php?script=sci_abstract&amp;pid=S0120-06902018000400285&amp;lng=en&amp;nrm=iso"></self-uri><self-uri xlink:href="http://www.scielo.org.co/scielo.php?script=sci_pdf&amp;pid=S0120-06902018000400285&amp;lng=en&amp;nrm=iso"></self-uri><abstract abstract-type="short" xml:lang="en"><p><![CDATA[Abstract  Background: Ehrlichia and Rickettsia are two major rickettsial genera transmitted by ticks that affect a number of wild and domestic animal species and human populations around the world.  Objective:  To design and validate a duplex PCR for Ehrlichia and Rickettsia in ticks.  Methods: Assay validation included testing for sensitivity, specificity, reproducibility, and robustness of the PCR. The groEL and 23sr RNA genes were used for Ehrlichia and Rickettsia, respectively.  Results: The limit of detection was one hundred gene copies per 50 &#956;L of reaction for Ehrlichia spp, and one gene copy of Rickettsia per 50 &#956;L of reaction. In general, the primers of the test only amplified in silico those bacterial agents for which they were originally designed, with the exception of the primers for Rickettsia that also amplified Methylocystis sp. The test was reproducible (intermediate precision) 96.7% of the times for both agents. The test was robust enough to tolerate concentration changes of all reagents with the exception of Taq DNA polymerase.  Conclusions: The validation results indicated that this PCR is useful for detection in both bacterial genera and it is a good candidate for diagnostic validation.]]></p></abstract>
<abstract abstract-type="short" xml:lang="es"><p><![CDATA[Resumen  Antecedentes: Ehrlichia spp. y Rickettsia spp. son dos de los principales géneros rickettsiales transmitidos por garrapatas que afectan a animales silvestres, domésticos y humanos alrededor del mundo.  Objetivo: Diseñar y validar una prueba PCR dúplex para Ehrlichia y Rickettsia en garrapatas.  Métodos: La validación de la prueba incluyó ensayos de sensibilidad, especificidad, reproducibilidad y robustez. En la PCR se usó groEL y ARNr 23S como genes blanco para Ehrlichia y Rickettsia, respectivamente.  Resultados: El límite de detección fue de 100 copias del gen por 50 &#956;L de reacción para Ehrlichia spp y una copia del gen de Rickettsia por 50 &#956;L de reacción. En general, los cebadores de la prueba solo amplificaron in silico los agentes bacterianos para los cuales fueron originalmente diseñados, con la excepción de los cebadores de Rickettsia que también amplificaron Methylocystis sp. La prueba fue reproducible (precisión intermedia) en un 96.7% de las veces para ambos agentes. La prueba fue suficientemente robusta como para tolerar cambios de concentración de los diferentes reactivos, con excepción de la Taq DNA polimerasa.  Conclusión: Los resultados de validación indican que la PCR es útil para detectar ambos géneros bacterianos y podria usarse para validación diagnostica.]]></p></abstract>
<abstract abstract-type="short" xml:lang="pt"><p><![CDATA[Resumo  Antecedentes: Ehrlichia e Rickettsia são dois dos principais gêneros de rickettsias transmitidos por carrapatos que infectam tanto animais selvagens quanto animais domésticos e até homens em todo o mundo.  Objetivo: O objetivo principal foi elaborar e validar uma PCR duplex para Ehrlichia e Rickettsia em carrapatos.  Métodos: A validação incluiu testes de sensibilidade, especificidade, reprodução e robustez. Para o PCR, utilizamos os genes groEl e 23Sr-RNA para Ehrlichia e Rickettsia, respectivamente.  Resultados:  O limite de detecção foi de 100 cópias de genes por 50 ml de reação para Erliquia spp e uma cópia de gene de Rickettsia por 50 ml de reação. Em geral, os iniciadores dos testes amplificaram em modelos computacionais os agentes bacterianos para os quais eles foram projetados, exceto os primers de Rickettsia que também amplificou Methylocystis sp. Os testes foram reproduzíveis (precisão intermediária) 96,7% para ambos os agentes e foram também robustos para tolerar mudanças de concentração em todos os reagentes, exceto o reagente Taq DNA polymerase.  Conclusões: Os resultados da validação indicaram que o PCR é útil para detecção em ambos os gêneros bacterianos, portanto, um bom exame para validação diagnóstica.]]></p></abstract>
<kwd-group>
<kwd lng="en"><![CDATA[exclusiveness]]></kwd>
<kwd lng="en"><![CDATA[groEL]]></kwd>
<kwd lng="en"><![CDATA[inclusiveness]]></kwd>
<kwd lng="en"><![CDATA[robustness]]></kwd>
<kwd lng="en"><![CDATA[rRNA 23S]]></kwd>
<kwd lng="es"><![CDATA[ARNr 23S]]></kwd>
<kwd lng="es"><![CDATA[exclusividad]]></kwd>
<kwd lng="es"><![CDATA[groEL]]></kwd>
<kwd lng="es"><![CDATA[inclusividad]]></kwd>
<kwd lng="es"><![CDATA[robustez]]></kwd>
<kwd lng="pt"><![CDATA[exclusividade]]></kwd>
<kwd lng="pt"><![CDATA[groEL]]></kwd>
<kwd lng="pt"><![CDATA[inclusividade]]></kwd>
<kwd lng="pt"><![CDATA[robustez]]></kwd>
<kwd lng="pt"><![CDATA[rRNA 23S]]></kwd>
</kwd-group>
</article-meta>
</front><back>
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