<?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>S0253-570X2021000300001</article-id>
<title-group>
<article-title xml:lang="en"><![CDATA[Polymerase chain reaction with internal amplification control to detect mycoplasmas in cell cultures and raw materials]]></article-title>
<article-title xml:lang="es"><![CDATA[Reacción en cadena de la polimerasa con control interno de amplificación para detectar micoplasmas en cultivos celulares y materias primas]]></article-title>
</title-group>
<contrib-group>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Burgher Pulgarón]]></surname>
<given-names><![CDATA[Yaima]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
<xref ref-type="aff" rid="Aaf"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Plasencia-Márquez]]></surname>
<given-names><![CDATA[Odaylin]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Pérez]]></surname>
<given-names><![CDATA[Anisleydis]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Peña]]></surname>
<given-names><![CDATA[Joan]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Corona-González]]></surname>
<given-names><![CDATA[Belkis]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Lobo-Rivero]]></surname>
<given-names><![CDATA[Evelyn]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
</contrib-group>
<aff id="Af1">
<institution><![CDATA[,National Center for Animal and Plant Health (CENSA) MYCOLAB (OIE Reference Laboratory for Mycoplasma Diagnosis) ]]></institution>
<addr-line><![CDATA[San José de las Lajas Mayabeque]]></addr-line>
<country>Cuba</country>
</aff>
<aff id="Af2">
<institution><![CDATA[,Université de Montréal Faculté de médecine vétérinaire The Swine and Poultry Infectious Diseases Research Centre (CRIPA) and the Research Group on Infectious Diseases in Production Animals (GREMIP)]]></institution>
<addr-line><![CDATA[ ]]></addr-line>
</aff>
<pub-date pub-type="pub">
<day>00</day>
<month>12</month>
<year>2021</year>
</pub-date>
<pub-date pub-type="epub">
<day>00</day>
<month>12</month>
<year>2021</year>
</pub-date>
<volume>43</volume>
<numero>3</numero>
<copyright-statement/>
<copyright-year/>
<self-uri xlink:href="http://scielo.sld.cu/scielo.php?script=sci_arttext&amp;pid=S0253-570X2021000300001&amp;lng=en&amp;nrm=iso"></self-uri><self-uri xlink:href="http://scielo.sld.cu/scielo.php?script=sci_abstract&amp;pid=S0253-570X2021000300001&amp;lng=en&amp;nrm=iso"></self-uri><self-uri xlink:href="http://scielo.sld.cu/scielo.php?script=sci_pdf&amp;pid=S0253-570X2021000300001&amp;lng=en&amp;nrm=iso"></self-uri><abstract abstract-type="short" xml:lang="en"><p><![CDATA[ABSTRACT The presence of polymerase chain reaction (PCR) inhibitory substances in cell cultures and raw materials used in the manufacture of monoclonal antibodies could limit the usefulness of this method to detect mycoplasma contamination. The objective of this research was to determine the sensitivity and specificity of a PCR method to detect mycoplasma deoxyribonucleic acid (DNA) in MDCK and HeLa cell lines, hybridoma cells, ascitic fluid and cell supernatant containing monoclonal antibodies. The effect of a thermal shock-based mycoplasma DNA extraction method on PCR sensitivity, as well as the performance of two mycoplasma DNA purification methods: silica/guanidinium thiocyanate and a commercial DNA purification kit, were evaluated. In addition, an internal amplification control (IAC) was optimized to detect inhibitory samples from these matrices after thermal shock treatment. PCR-IAC was specific for the amplification of mycoplasma DNA. The inclusion of IAC plasmid at the concentration selected did not decrease the sensitivity of PCR in mycoplasma-spiked matrices. Seventy-eight samples were analyzed; all represented the matrices under study. As a result, all matrices showed PCR inhibition after thermal shock. The inhibitory effect decreased when silica/guanidinium thiocyanate or commercial DNA purification kit was used. PCR-IAC detected 65 % of inhibitory samples and revealed differential susceptibility to inhibitors among different samples of the same matrix.]]></p></abstract>
<abstract abstract-type="short" xml:lang="es"><p><![CDATA[RESUMEN La presencia de sustancias inhibitorias de la reacción en cadena de la polimerasa (PCR) en cultivos celulares y materias primas usadas en la fabricación de anticuerpos monoclonales podría limitar la utilidad de esta técnica para detectar la contaminación por micoplasmas. El objetivo de esta investigación fue determinar la sensibilidad y la especificidad de un método de PCR para detectar ácido desorribunucleico (ADN) de micoplasma en las líneas celulares MDCK y HELA, en células de hibridoma, en líquido ascítico y en sobrenadante celular que contenía un anticuerpo monoclonal. Se evaluaron el efecto de un método de extracción de ADN micoplásmico, basado en choque térmico sobre la sensibilidad de la PCR, así como el desempeño de dos métodos de purificación de ADN micoplásmico: sílice / tiocianato de guanidinio y un kit comercial de purificación de ADN. Además, se optimizó un control de amplificación interno (CAI) para detectar muestras inhibitorias de estas matrices después del tratamiento de choque térmico. La PCR-CAI fue específica para la amplificación del ADN de micoplasma; la inclusión del plásmido CAI a la concentración seleccionada no disminuyó la sensibilidad de la PCR en las matrices experimentalmente contaminadas con micoplasma. Se analizaron 78 muestras; todas representaron las matrices en estudio. Como resultado, todas las matrices mostraron inhibición de la PCR después del choque térmico. El efecto inhibitorio disminuyó cuando se utilizó sílice / tiocianato de guanidinio o el kit comercial de purificación de ADN. La PCR-CAI detectó 65 % de muestras inhibitorias y puso en evidencia una susceptibilidad diferencial a los inhibidores entre diferentes muestras de una misma matriz.]]></p></abstract>
<kwd-group>
<kwd lng="en"><![CDATA[PCR]]></kwd>
<kwd lng="en"><![CDATA[inhibition]]></kwd>
<kwd lng="en"><![CDATA[internal amplification control]]></kwd>
<kwd lng="en"><![CDATA[monoclonal antibodies]]></kwd>
<kwd lng="en"><![CDATA[mycoplasma]]></kwd>
<kwd lng="es"><![CDATA[PCR]]></kwd>
<kwd lng="es"><![CDATA[inhibición]]></kwd>
<kwd lng="es"><![CDATA[control de amplificación interno]]></kwd>
<kwd lng="es"><![CDATA[anticuerpos monoclonales]]></kwd>
<kwd lng="es"><![CDATA[micoplasma]]></kwd>
</kwd-group>
</article-meta>
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