<?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>2176-6223</journal-id>
<journal-title><![CDATA[Revista Pan-Amazônica de Saúde]]></journal-title>
<abbrev-journal-title><![CDATA[Rev Pan-Amaz Saude]]></abbrev-journal-title>
<issn>2176-6223</issn>
<publisher>
<publisher-name><![CDATA[Instituto Evandro Chagas. Secretaria de Vigilância em Saúde e Ambiente. Ministério da Saúde]]></publisher-name>
</publisher>
</journal-meta>
<article-meta>
<article-id>S2176-62232016000500107</article-id>
<article-id pub-id-type="doi">10.5123/s2176-62232016000500012</article-id>
<title-group>
<article-title xml:lang="en"><![CDATA[Nested PCR using thick blood smears as source of Plasmodium DNA: an alternative to study archival blood films]]></article-title>
<article-title xml:lang="pt"><![CDATA[Nested PCR utilizando a gota espessa como fonte do DNA de Plasmodium: uma alternativa para análise de coleções de lâminas de sangue arquivadas]]></article-title>
<article-title xml:lang="es"><![CDATA[Nested PCR utilizando la gota gruesa como fuente del ADN de Plasmodium: una alternativa para f rotis de sangre archivados]]></article-title>
</title-group>
<contrib-group>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Viana]]></surname>
<given-names><![CDATA[Giselle Maria Rachid]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Chamma-Siqueira]]></surname>
<given-names><![CDATA[Nathália Nogueira]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Barbosa]]></surname>
<given-names><![CDATA[Danielle Regina Lima]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Carmo]]></surname>
<given-names><![CDATA[Ediclei Lima do]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Peres]]></surname>
<given-names><![CDATA[José Mário Veloso]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Nascimento]]></surname>
<given-names><![CDATA[José Maria de Souza]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Póvoa]]></surname>
<given-names><![CDATA[Marinete Marins]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
</contrib-group>
<aff id="Af1">
<institution><![CDATA[,Instituto Evandro Chagas/SVS/MS Seção de Parasitologia ]]></institution>
<addr-line><![CDATA[Ananindeua Pará]]></addr-line>
<country>Brasil</country>
</aff>
<pub-date pub-type="pub">
<day>00</day>
<month>12</month>
<year>2016</year>
</pub-date>
<pub-date pub-type="epub">
<day>00</day>
<month>12</month>
<year>2016</year>
</pub-date>
<volume>7</volume>
<numero>esp</numero>
<fpage>107</fpage>
<lpage>114</lpage>
<copyright-statement/>
<copyright-year/>
<self-uri xlink:href="http://scielo.iec.gov.br/scielo.php?script=sci_arttext&amp;pid=S2176-62232016000500107&amp;lng=en&amp;nrm=iso"></self-uri><self-uri xlink:href="http://scielo.iec.gov.br/scielo.php?script=sci_abstract&amp;pid=S2176-62232016000500107&amp;lng=en&amp;nrm=iso"></self-uri><self-uri xlink:href="http://scielo.iec.gov.br/scielo.php?script=sci_pdf&amp;pid=S2176-62232016000500107&amp;lng=en&amp;nrm=iso"></self-uri><abstract abstract-type="short" xml:lang="en"><p><![CDATA[ABSTRACT The study aimed to evaluate a protocol of nested PCR using archival Giemsa-stained thick blood smears (GTS)as source of Plasmodium DNA. A total of 138 GTS from patients of five municipalities from Pará State (Amazon Region, Brazil) was included in this survey. These samples were classified in three groups (group 1: 85 Plasmodium positive and negative GTS stored in plastic box during five years; group 2: 28 Plasmodium positive and negative GTS stored in wooden box during 10 years; and group 3: 25 Trypanosoma cruzi GTS negative for Plasmodium stored in plastic box during a month) and were submitted to DNA extraction with Chelex-100. Subsequently, extracted DNA samples were quantified and the integrity was verified by electrophoresis. Nested PCR protocol was performed to detect Plasmodium species. The results of nested PCR were compared to microscopy and statistic parameters were calculated by screening test. DNA samples from all groups had acceptable quantity and purity level, but the evaluation of integrity showed 19 degraded samples from group 2. By nested PCR, this group showed very low sensitivity (29.63%) and accuracy (32.14%), while nested PCR for samples from group 1 showed 100% of sensitivity and 97.65% of accuracy. The results of this research showed that samples stored until five years can be useful as Plasmodium DNA source for nested PCR to identify Plasmodium species, being an important alternative to support retrospective studies.]]></p></abstract>
<abstract abstract-type="short" xml:lang="pt"><p><![CDATA[RESUMO O estudo teve como objetivo avaliar um protocolo de nested PCR, utilizando a gota espessa (GE) corada por Giemsa como fonte de DNA de Plasmodium. Um total de 138 amostras de GE de pacientes de cinco municípios do Estado do Pará (Região Amazônica, Brasil) foi incluído neste estudo. Essas amostras foram classificadas em três grupos (grupo 1: 85 lâminas de GE positivas e negativas para Plasmodium armazenadas em caixa plástica por cinco anos; grupo 2: 28 lâminas de GE positivas e negativas para Plasmodium armazenadas em caixa de madeira por 10 anos; e grupo 3: 25 lâminas de GE positivas para Trypanosoma cruzi e negativas para Plasmodium) e submetidas à extração de DNA com Chelex-100. Posteriormente, as amostras de DNA extraídas foram quantificadas e a integridade verificada por eletroforese. O protocolo de nested PCR foi realizado para detectar as espécies de Plasmodium. Os resultados foram então comparados com os da microscopia e os parâmetros estatísticos calculados pelo screening test. As amostras de DNA de todos os grupos apresentaram quantidade e níveis de pureza adequados para realização de PCR, mas a avaliação da integridade mostrou 19 amostras degradadas do grupo 2. Pelo nested PCR, esse grupo mostrou baixa sensibilidade (29,63%) e acurácia (32,14%), enquanto que as amostras do grupo 1 apresentaram 100% de sensibilidade e 97,65% de acurácia. Os resultados desta pesquisa apontam que as amostras armazenadas até cinco anos podem ser úteis como fonte de DNA de Plasmodium para que o nested PCR identifique as espécies desse gênero, sendo uma alternativa importante para apoiar estudos retrospectivos.]]></p></abstract>
<abstract abstract-type="short" xml:lang="es"><p><![CDATA[RESUMEN El estudio tuvo como objetivo evaluar un protocolo de nested PCR, utilizando la prueba de la gota gruesa teñida por Giemsa (GG) como fuente de ADN de Plasmodium. Un total de 138 muestras de GG de pacientes de cinco municipios del Estado de Pará (Región Amazónica, Brasil) se incluyó en este estudio; estas muestras fueron clasificadas en tres grupos (grupo 1: 85 láminas de GE positivas y negativas para Plasmodium almacenados en una caja de plástico durante cinco años; grupo 2: 28 láminas de GG positivas y negativas para Plasmodium almacenados en una caja de madera durante diez años; y grupo 3: 25 láminas de GG positivas para Trypanosoma cruzi y negativas para Plasmodium, y sometidas a la extracción de ADN con Chelex-100. Posteriormente las muestras de ADN extraídas se cuantificaron y su integridad se verificó por electroforesis. El protocolo de nested PCR fue realizado para detectar las especies de Plasmodium. Los resultados de nested PCR fueron comparados con los de la microscopia y los parámetros estadísticos se calcularon por la prueba de selección. Las muestras de ADN de todos los grupos obtuvieron nivel de cantidad y de pureza adecuadas para la realización de PCR, pero la evaluación de integridad mostró 19 muestras degradadas del grupo 2. Por nested PCR, este grupo mostró baja sensibilidad (29,63%) y precisión (32,14%), mientras que las muestras del grupo 1 presentaron 100% de sensibilidad y 97,65% de precisión. Los resultados de esta investigación señalan que las muestras almacenadas por hasta cinco años pueden ser útiles como fuente de ADN de Plasmodium para que nested PCR identifique las especies de ese género, siendo una alternativa importante para apoyar estudios retrospectivos.]]></p></abstract>
<kwd-group>
<kwd lng="en"><![CDATA[Malaria]]></kwd>
<kwd lng="en"><![CDATA[Thick Blood Smears]]></kwd>
<kwd lng="en"><![CDATA[Polymerase Chain Reaction]]></kwd>
<kwd lng="en"><![CDATA[Diagnostic Techniques and Procedures]]></kwd>
<kwd lng="pt"><![CDATA[Malária]]></kwd>
<kwd lng="pt"><![CDATA[Gota Espessa]]></kwd>
<kwd lng="pt"><![CDATA[Reação em Cadeia da Polimerase]]></kwd>
<kwd lng="pt"><![CDATA[Técnicas e Procedimentos em Diagnóstico.]]></kwd>
<kwd lng="es"><![CDATA[Malaria]]></kwd>
<kwd lng="es"><![CDATA[Gota gruesa]]></kwd>
<kwd lng="es"><![CDATA[Reacción en Cadena de la Polimerasa]]></kwd>
<kwd lng="es"><![CDATA[Técnicas y Procedimientos en Diagnóstico]]></kwd>
</kwd-group>
</article-meta>
</front><back>
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