<?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>1887-8571</journal-id>
<journal-title><![CDATA[Sanidad Militar]]></journal-title>
<abbrev-journal-title><![CDATA[Sanid. Mil.]]></abbrev-journal-title>
<issn>1887-8571</issn>
<publisher>
<publisher-name><![CDATA[Ministerio de Defensa]]></publisher-name>
</publisher>
</journal-meta>
<article-meta>
<article-id>S1887-85712018000200084</article-id>
<article-id pub-id-type="doi">10.4321/s1887-85712018000200004</article-id>
<title-group>
<article-title xml:lang="es"><![CDATA[Optimización y Validación de una PCR en Tiempo Real para la Rápida Identificación de Bacillus thuringiensis, Simulador de Bacillus anthracis]]></article-title>
<article-title xml:lang="en"><![CDATA[Optimization and validation of a real-time PCR for rapid identification of Bacillus thuringiensis, surrogate of Bacillus anthracis]]></article-title>
</title-group>
<contrib-group>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Bassy Álvarez]]></surname>
<given-names><![CDATA[O]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
<xref ref-type="aff" rid="Aaf"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Larigauderie]]></surname>
<given-names><![CDATA[G]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Ortega García]]></surname>
<given-names><![CDATA[MªV]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Granja Albarellos]]></surname>
<given-names><![CDATA[C]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Cabria Ramos]]></surname>
<given-names><![CDATA[JC]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
</contrib-group>
<aff id="Af1">
<institution><![CDATA[,IMIDEF INTA Dpto. de Sistemas de Defensa NBQ y Materiales Energéticos, Área de Defensa Biológica, Campus La Marañosa]]></institution>
<addr-line><![CDATA[Madrid ]]></addr-line>
<country>España</country>
</aff>
<aff id="Af6">
<institution><![CDATA[,ISDEFE  ]]></institution>
<addr-line><![CDATA[Madrid ]]></addr-line>
<country>España</country>
</aff>
<aff id="Af2">
<institution><![CDATA[,DGA Maîtrise NRBC  ]]></institution>
<addr-line><![CDATA[Vert-le-Petit ]]></addr-line>
<country>Francia</country>
</aff>
<aff id="Af3">
<institution><![CDATA[,IMIDEF INTA Dpto. de Sistemas de Defensa NBQ y Materiales Energéticos, Área de Defensa Biológica, Campus La Marañosa]]></institution>
<addr-line><![CDATA[Madrid ]]></addr-line>
<country>España</country>
</aff>
<aff id="Af4">
<institution><![CDATA[,IMIDEF INTA Dpto. de Sistemas de Defensa NBQ y Materiales Energéticos, Área de Defensa Biológica, Campus La Marañosa]]></institution>
<addr-line><![CDATA[Madrid ]]></addr-line>
<country>España</country>
</aff>
<aff id="Af5">
<institution><![CDATA[,IMIDEF INTA Dpto. de Sistemas de Defensa NBQ y Materiales Energéticos, Área de Defensa Biológica, Campus La Marañosa]]></institution>
<addr-line><![CDATA[Madrid ]]></addr-line>
<country>España</country>
</aff>
<pub-date pub-type="pub">
<day>00</day>
<month>06</month>
<year>2018</year>
</pub-date>
<pub-date pub-type="epub">
<day>00</day>
<month>06</month>
<year>2018</year>
</pub-date>
<volume>74</volume>
<numero>2</numero>
<fpage>84</fpage>
<lpage>89</lpage>
<copyright-statement/>
<copyright-year/>
<self-uri xlink:href="http://scielo.isciii.es/scielo.php?script=sci_arttext&amp;pid=S1887-85712018000200084&amp;lng=en&amp;nrm=iso"></self-uri><self-uri xlink:href="http://scielo.isciii.es/scielo.php?script=sci_abstract&amp;pid=S1887-85712018000200084&amp;lng=en&amp;nrm=iso"></self-uri><self-uri xlink:href="http://scielo.isciii.es/scielo.php?script=sci_pdf&amp;pid=S1887-85712018000200084&amp;lng=en&amp;nrm=iso"></self-uri><abstract abstract-type="short" xml:lang="es"><p><![CDATA[RESUMEN  Introducción: Bacillus anthracis es uno de los agentes de guerra biológica más empleados en el mundo. Sin embargo, en los laboratorios de defensa biológica es conveniente utilizar en muchas ocasiones otras bacterias parecidas a Bacillus anthracis pero menos peligrosas o apatógenas. Uno de los principales simuladores de Bacillus anthracis es Bacillus thuringiensis, debido a su elevada homología con B. anthracis y a su nula patogenicidad para el ser humano.  Objetivos: El objetivo del presente estudio es desarrollar y validar una PCR en tiempo real para la rápida identificación del ADN de Bacillus thuringiensis, agente empleado muy a menudo en los simulacros para entrenamiento de las Unidades Operativas de toma de muestras NBQ de las FAS.  Material y Métodos: La identificación del simulador Bacillus thuringiensis se ha realizado mediante la amplificación y detección con una sonda de hidrólisis de un fragmento de 69 pares de bases del gen cry1A, el cual es específico de esta bacteria. Tras optimizar las condiciones de amplificación probando tres temperaturas diferentes de hibridación/extensión, se procedió a realizar la validación del método desarrollado.  Resultados: La nueva PCR en tiempo real desarrollada presentó una eficiencia del 93%, así como una elevada linealidad (coeficiente de regresión R2 0,9993). El límite de detección al 95% de probabilidad fue de 13 equivalentes de genoma completo por reacción. Tanto la inclusividad como la exclusividad del método fueron del 100%.  Conclusiones: El método molecular desarrollado por el Laboratorio de Biología Molecular del INTA permite una rápida identificación del ADN de Bacillus thuringiensis, simulador de Bacillus anthracis, con unas elevadas sensibilidad y especificidad analíticas.]]></p></abstract>
<abstract abstract-type="short" xml:lang="en"><p><![CDATA[SUMMARY  Introduction: Bacillus anthracis is the most employed biological warfare agent in the world. However, in biological defense laboratories, many times it is convenient to use other bacteria similar to Bacillus anthracis but less dangerous or non-pathogenic. One of the main surrogates of Bacillus anthracis is Bacillus thuringiensis, due to its high homology with B. anthracis and its null pathogenicity for humans.  Objective: The objective of the present study is to develope and validate a real-time PCR for the rapid identification of Bacillus thuringiensis DNA, biological agent very often employed in the training of the Operative Units of CBRN sampling of the Armed Forces.  Methods: The identification of Bacillus thuringiensis has been performed by the amplification and detection with a hydrolysis probe of a 69 base pairs fragment of the cry1A gene, which is specific for this bacterium. After optimizing the amplification conditions by testing three different hybridization / extension temperatures, the validation of the new developed method was carried out.  Results: The new developed real-time PCR showed an efficiency of 93%, as well as a high linearity (regression coefficient R20.9993). The limit of detection at 95% probability was 13 genome equivalents per reaction. Both the inclusiveness and the exclusivity of the method were 100%.  Conclusions: The molecular method developed at the Molecular Biology Laboratory of INTA allows the rapid identification of Bacillus thuringiensis DNA, surrogate of Bacillus anthracis, with a high analytical sensitivity and specificity.]]></p></abstract>
<kwd-group>
<kwd lng="es"><![CDATA[Bacillus thuringiensis]]></kwd>
<kwd lng="es"><![CDATA[PCR en tiempo real]]></kwd>
<kwd lng="es"><![CDATA[identificación molecular]]></kwd>
<kwd lng="es"><![CDATA[gen cry1A]]></kwd>
<kwd lng="en"><![CDATA[Bacillus thuringiensis]]></kwd>
<kwd lng="en"><![CDATA[real-time PCR]]></kwd>
<kwd lng="en"><![CDATA[molecular identification]]></kwd>
<kwd lng="en"><![CDATA[cry1A gen]]></kwd>
</kwd-group>
</article-meta>
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