<?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>0121-0807</journal-id>
<journal-title><![CDATA[Revista de la Universidad Industrial de Santander. Salud]]></journal-title>
<abbrev-journal-title><![CDATA[Rev. Univ. Ind. Santander. Salud]]></abbrev-journal-title>
<issn>0121-0807</issn>
<publisher>
<publisher-name><![CDATA[Universidad Industrial de Santander]]></publisher-name>
</publisher>
</journal-meta>
<article-meta>
<article-id>S0121-08072025000100036</article-id>
<article-id pub-id-type="doi">10.18273/saluduis.57.e:25v58a33</article-id>
<title-group>
<article-title xml:lang="en"><![CDATA[Tetramethylammonium Chloride and Dimethyl Sulfoxide improve SARS-CoV-2 detection in individual/pooled saliva samples with RT-qPCR]]></article-title>
<article-title xml:lang="es"><![CDATA[Cloruro de Tetrametilamonio y Dimetilsulfóxido mejoran la detección de SARS-CoV-2 en muestras de saliva individuales/agrupadas con RT-qPCR]]></article-title>
</title-group>
<contrib-group>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Vera-Cala]]></surname>
<given-names><![CDATA[Lina María]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Navarro-Baron]]></surname>
<given-names><![CDATA[Nathaniel A.]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Cangrejo-Useda]]></surname>
<given-names><![CDATA[Yordy S.]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Pardo-Diaz]]></surname>
<given-names><![CDATA[Luis A.]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Cadena-Caballero]]></surname>
<given-names><![CDATA[Cristian E.]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Barrios Hernández]]></surname>
<given-names><![CDATA[Carlos J.]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
<xref ref-type="aff" rid="Aaf"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Martinez-Perez]]></surname>
<given-names><![CDATA[Francisco]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
</contrib-group>
<aff id="Af1">
<institution><![CDATA[,Universidad Industrial de Santander  ]]></institution>
<addr-line><![CDATA[Bucaramanga Santander]]></addr-line>
<country>Colombia</country>
</aff>
<aff id="Af2">
<institution><![CDATA[,Instituto Nacional de Investigación en Informática y Automática  ]]></institution>
<addr-line><![CDATA[Grenoble ]]></addr-line>
<country>Francia</country>
</aff>
<pub-date pub-type="pub">
<day>00</day>
<month>12</month>
<year>2025</year>
</pub-date>
<pub-date pub-type="epub">
<day>00</day>
<month>12</month>
<year>2025</year>
</pub-date>
<volume>57</volume>
<copyright-statement/>
<copyright-year/>
<self-uri xlink:href="http://www.scielo.org.co/scielo.php?script=sci_arttext&amp;pid=S0121-08072025000100036&amp;lng=en&amp;nrm=iso"></self-uri><self-uri xlink:href="http://www.scielo.org.co/scielo.php?script=sci_abstract&amp;pid=S0121-08072025000100036&amp;lng=en&amp;nrm=iso"></self-uri><self-uri xlink:href="http://www.scielo.org.co/scielo.php?script=sci_pdf&amp;pid=S0121-08072025000100036&amp;lng=en&amp;nrm=iso"></self-uri><abstract abstract-type="short" xml:lang="en"><p><![CDATA[Abstract  Introduction: RT-qPCR performance to detect SARS-CoV-2 varies because SNPs at primer/probe sites and RNA secondary structures (notably 5'/3' UTR stem-loops) can hinder hybridization/polymerase elongation, altering Ct and fluorescence. A denaturant solution (DS: composed of Tetramethylammonium Chloride and Dimethyl Sulfoxide) can disrupt these structures and improves its detection.  Objective: Evaluate the DS efficiency in RT-qPCR to SARS-CoV-2 detection from single and multiplex saliva samples in COVID-19 positive patients.  Methodology: To establish the hybridization pattern, in silico analysis was carried out among primers and probes authorized by CDC (Centers for Disease Control and Prevention) with respect to the consensus region of SARS-CoV-2 N gen, and their secondary and tertiary structures were generated to establish variable regions. Individual and pooled RT-qPCR with DS reactions, were validated from total RNA from 20 of 40 saliva samples positive to SARS-CoV-2.  Results: N1 consensus region of SARS-CoV-2 variants: Alfa, Beta, Delta, Gamma, GH490R Lambda, Mu y Ómicron from 2021, exhibited a high variation respect to N2 region, but were located in bubbles near to a stem and in the spacer region. Individual RT-qPCR to N1 and N2 regions showed no significant statistical differences in Cts, however, they were lower, with a remarkable difference in fluorescence signal, to N1 region in pooled RT-qPCR reactions.  Conclusion: The procedure evidences the advantages in the inclusion of DS in RT-qPCR to SARS-CoV-2 detection in saliva samples.]]></p></abstract>
<abstract abstract-type="short" xml:lang="es"><p><![CDATA[Resumen  Introducción: El desempeño de la RT-qPCR para detectar SARS-CoV-2 varía porque los SNP en los sitios de cebadores/ sondas y las estructuras secundarias del ARN (en particular, los bucles de tallo en las UTR 5'/3') pueden obstaculizar la hibridación y la elongación de la polimerasa, alterando el Ct y la fluorescencia. Una solución desnaturalizante (DS; compuesta por cloruro de tetrametilamonio y dimetilsulfóxido) puede alterar estas estructuras y mejorar la detección.  Objetivo: Evaluar el efecto de la SD en la eficiencia de la RT-qPCR para la detección de SARS-CoV-2 en muestras de saliva individuales o agrupadas de pacientes diagnosticados con COVID-19.  Metodología: Se determinó in silico el patrón de hibridación entre los cebadores y sondas autorizados por el CDC (Centers for Disease Control and Prevention) respecto a la región consenso del gen N de SARS-CoV-2, además de generar su estructura secundaria para definir las posiciones variables. La validación de las RT-qPCR individuales y agrupadas con la SD se realizó con ARN total en 20 de 40 muestras de saliva positivas a SARS-CoV-2.  Resultados: La región consenso del gen N1 de las variantes de SARS-CoV-2: Alfa, Beta, Delta, Gamma, GH490R, Lambda, Mu y Ómicron del año 2021, presentó más variaciones respecto a la región N2, no obstante, se localizaron dentro de las burbujas adyacentes a un tallo y en la región espaciadora. Las RT-qPCR individuales para las regiones N1 y N2 con SD no mostraron Cts estadísticamente significativos, sin embargo, estos disminuyeron, con una diferencia notable en la señal de fluorescencia, para la región N1 en las reacciones de RT-qPCR agrupadas.  Conclusión: El procedimiento confirma la ventaja de la inclusión de la SD en la RT-qPCR para detectar SARS-CoV-2 en muestras de saliva.]]></p></abstract>
<kwd-group>
<kwd lng="en"><![CDATA[SARS-CoV-2]]></kwd>
<kwd lng="en"><![CDATA[Saliva Samples]]></kwd>
<kwd lng="en"><![CDATA[RT-qPCR Multiplex]]></kwd>
<kwd lng="en"><![CDATA[Coadjuvant Solution]]></kwd>
<kwd lng="en"><![CDATA[RNA Secondary Structure]]></kwd>
<kwd lng="es"><![CDATA[SARS-CoV-2]]></kwd>
<kwd lng="es"><![CDATA[Muestras de Saliva]]></kwd>
<kwd lng="es"><![CDATA[RT-qPCR múltiples]]></kwd>
<kwd lng="es"><![CDATA[Solución Coadyuvante]]></kwd>
<kwd lng="es"><![CDATA[Estructura Secundaria del ARN]]></kwd>
</kwd-group>
</article-meta>
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