Otimização da Reação em Cadeia da Polimerase (PCR) para diferenciação molecular de amebas do complexo Histolytica
Main Author: | |
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Publication Date: | 2018 |
Format: | Bachelor thesis |
Language: | por |
Source: | Repositório Institucional da UFS |
Download full: | http://ri.ufs.br/jspui/handle/riufs/9595 |
Summary: | Intestinal parasites contribute to increased morbidity and mortality rates in the world, with amebiasis being the second leading cause of death among parasitic diseases, with approximately 100,000 deaths per year. To prevent unnecessary treatment of individuals with commensal Entamoeba species, it is extremely important to distinguish them from potentially pathogenic E. histolytica. Optical microscopic examination of feces is not efficient for the differentiation of Entamoeba species, and molecular methods are considered useful and reliable for the identification and differentiation of these species, including conventional PCR, Nested-PCR and Multiplex-PCR. The aim of this study was to optimize PCR techniques for better results in the differentiation of amoeba species from the histolytic complex and Entamoeba hartmanni. For this, 16 positive stool samples were obtained by optical microscopy for Entamoeba histolytica/E. dispar. The results of the present study were similar to those of E. hartmanni, Nested-PCR, and Multiplex-PCR for E. histolytica, E. dispar and E. moshkovskii, with modifications of the original protocols to obtain better results in its standardization. In Conventional PCR two positive samples were observed for E. hartmanni, in the Nested-PCR one sample was positive for E. dispar and for E. moshkovskii species no sample was positive. Although the results of the optimization were not completely satisfactory in all the samples tested, due to the possible degradation of the reagents used, the previous results obtained demonstrate the importance of PCR as an effective tool for the detection and differentiation of species of the genus Entamoeba |
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Mesquita, Danielle Marcela Menezes deDolabella, Silvio Santana2018-11-07T13:56:18Z2018-11-07T13:56:18Z2018-10-25Mesquita, Danielle Marcela Menezes de. Otimização da Reação em Cadeia da Polimerase (PCR) para diferenciação molecular de amebas do complexo Histolytica. São Cristóvão, SE, 2018. Monografia (graduação de Biologia) – Departamento de Biologia, Centro de Ciências Biológicas e da Saúde, Universidade Federal de Sergipe, São Cristóvão, 2018http://ri.ufs.br/jspui/handle/riufs/9595Intestinal parasites contribute to increased morbidity and mortality rates in the world, with amebiasis being the second leading cause of death among parasitic diseases, with approximately 100,000 deaths per year. To prevent unnecessary treatment of individuals with commensal Entamoeba species, it is extremely important to distinguish them from potentially pathogenic E. histolytica. Optical microscopic examination of feces is not efficient for the differentiation of Entamoeba species, and molecular methods are considered useful and reliable for the identification and differentiation of these species, including conventional PCR, Nested-PCR and Multiplex-PCR. The aim of this study was to optimize PCR techniques for better results in the differentiation of amoeba species from the histolytic complex and Entamoeba hartmanni. For this, 16 positive stool samples were obtained by optical microscopy for Entamoeba histolytica/E. dispar. The results of the present study were similar to those of E. hartmanni, Nested-PCR, and Multiplex-PCR for E. histolytica, E. dispar and E. moshkovskii, with modifications of the original protocols to obtain better results in its standardization. In Conventional PCR two positive samples were observed for E. hartmanni, in the Nested-PCR one sample was positive for E. dispar and for E. moshkovskii species no sample was positive. Although the results of the optimization were not completely satisfactory in all the samples tested, due to the possible degradation of the reagents used, the previous results obtained demonstrate the importance of PCR as an effective tool for the detection and differentiation of species of the genus EntamoebaAs parasitoses intestinais contribuem para o aumento das taxas de morbidade e mortalidade no mundo, sendo a amebíase a segunda causa de morte entre as doenças parasitárias, com aproximadamente 100.000 mortes por ano. Para impedir que ocorra tratamento desnecessário de indivíduos com espécies de Entamoeba comensais, é de extrema importância diferenciá-las da Entamoeba histolytica, potencialmente patogênica. O exame microscópico de amostras fecais não é eficiente para a diferenciação das espécies de Entamoeba, sendo os métodos moleculares considerados úteis e confiáveis para a identificação e diferenciação dessas espécies, incluindo a PCR convencional, Nested-PCR e a Multiplex-PCR. O objetivo desse estudo foi otimizar técnicas de PCR para uma melhor obtenção de resultados na diferenciação de espécies de amebas do complexo histolytica e da Entamoeba hartmanni. Para isso, 16 amostras de fezes positivas na microscopia óptica para E. histolytica/E. dispar tiveram seu DNA extraído e, posteriormente, foram realizadas a PCR convencional para E. hartmanni, a Nested-PCR e Multiplex-PCR para E. histolytica, E. dispar e E. moshkovskii, com modificações dos protocolos originais para obtenção de melhores resultados na sua padronização. Na PCR Convencional foram observadas duas amostras positivas para E. hartmanni, no Nested-PCR uma amostra foi positiva para E. dispar e para a espécie E. moshkovskii nenhuma amostra foi positiva. Embora os resultados da otimização não tenham sido completamente satisfatórios em todas as amostras testadas devido à possível degradação dos reativos utilizados, os resultados prévios obtidos demonstram a importância da PCR como ferramenta eficaz para a detecção e diferenciação de espécies do gênero EntamoebaSão Cristóvão, SEporBiologiaEnsino de biologiaAmebíaseDiagnóstico molecularMétodos molecularesAmebiasisMolecular methodsCIENCIAS BIOLOGICAS::BIOQUIMICA::BIOLOGIA MOLECULAROtimização da Reação em Cadeia da Polimerase (PCR) para diferenciação molecular de amebas do complexo Histolyticainfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/bachelorThesisUniversidade Federal de SergipeDBI - Departamento de Biologia – São Cristóvão – Presencialreponame:Repositório Institucional da UFSinstname:Universidade Federal de Sergipe (UFS)instacron:UFSinfo:eu-repo/semantics/openAccessTEXTDanielle_Marcela_Menezes_Mesquita.pdf.txtDanielle_Marcela_Menezes_Mesquita.pdf.txtExtracted texttext/plain51990https://ri.ufs.br/jspui/bitstream/riufs/9595/3/Danielle_Marcela_Menezes_Mesquita.pdf.txt0957c968c9c502d1da61c350a75b1198MD53THUMBNAILDanielle_Marcela_Menezes_Mesquita.pdf.jpgDanielle_Marcela_Menezes_Mesquita.pdf.jpgGenerated Thumbnailimage/jpeg1281https://ri.ufs.br/jspui/bitstream/riufs/9595/4/Danielle_Marcela_Menezes_Mesquita.pdf.jpg55e050389198df69c8f328bf996a3f09MD54LICENSElicense.txtlicense.txttext/plain; charset=utf-81475https://ri.ufs.br/jspui/bitstream/riufs/9595/1/license.txt098cbbf65c2c15e1fb2e49c5d306a44cMD51ORIGINALDanielle_Marcela_Menezes_Mesquita.pdfDanielle_Marcela_Menezes_Mesquita.pdfapplication/pdf1294987https://ri.ufs.br/jspui/bitstream/riufs/9595/2/Danielle_Marcela_Menezes_Mesquita.pdfc16f51838efaba0aee2cac6f4a566982MD52riufs/95952018-11-07 10:56:18.688oai:ufs.br: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Repositório InstitucionalPUBhttps://ri.ufs.br/oai/requestrepositorio@academico.ufs.bropendoar:2018-11-07T13:56:18Repositório Institucional da UFS - Universidade Federal de Sergipe (UFS)false |
dc.title.pt_BR.fl_str_mv |
Otimização da Reação em Cadeia da Polimerase (PCR) para diferenciação molecular de amebas do complexo Histolytica |
title |
Otimização da Reação em Cadeia da Polimerase (PCR) para diferenciação molecular de amebas do complexo Histolytica |
spellingShingle |
Otimização da Reação em Cadeia da Polimerase (PCR) para diferenciação molecular de amebas do complexo Histolytica Mesquita, Danielle Marcela Menezes de Biologia Ensino de biologia Amebíase Diagnóstico molecular Métodos moleculares Amebiasis Molecular methods CIENCIAS BIOLOGICAS::BIOQUIMICA::BIOLOGIA MOLECULAR |
title_short |
Otimização da Reação em Cadeia da Polimerase (PCR) para diferenciação molecular de amebas do complexo Histolytica |
title_full |
Otimização da Reação em Cadeia da Polimerase (PCR) para diferenciação molecular de amebas do complexo Histolytica |
title_fullStr |
Otimização da Reação em Cadeia da Polimerase (PCR) para diferenciação molecular de amebas do complexo Histolytica |
title_full_unstemmed |
Otimização da Reação em Cadeia da Polimerase (PCR) para diferenciação molecular de amebas do complexo Histolytica |
title_sort |
Otimização da Reação em Cadeia da Polimerase (PCR) para diferenciação molecular de amebas do complexo Histolytica |
author |
Mesquita, Danielle Marcela Menezes de |
author_facet |
Mesquita, Danielle Marcela Menezes de |
author_role |
author |
dc.contributor.author.fl_str_mv |
Mesquita, Danielle Marcela Menezes de |
dc.contributor.advisor1.fl_str_mv |
Dolabella, Silvio Santana |
contributor_str_mv |
Dolabella, Silvio Santana |
dc.subject.por.fl_str_mv |
Biologia Ensino de biologia Amebíase Diagnóstico molecular Métodos moleculares |
topic |
Biologia Ensino de biologia Amebíase Diagnóstico molecular Métodos moleculares Amebiasis Molecular methods CIENCIAS BIOLOGICAS::BIOQUIMICA::BIOLOGIA MOLECULAR |
dc.subject.eng.fl_str_mv |
Amebiasis Molecular methods |
dc.subject.cnpq.fl_str_mv |
CIENCIAS BIOLOGICAS::BIOQUIMICA::BIOLOGIA MOLECULAR |
description |
Intestinal parasites contribute to increased morbidity and mortality rates in the world, with amebiasis being the second leading cause of death among parasitic diseases, with approximately 100,000 deaths per year. To prevent unnecessary treatment of individuals with commensal Entamoeba species, it is extremely important to distinguish them from potentially pathogenic E. histolytica. Optical microscopic examination of feces is not efficient for the differentiation of Entamoeba species, and molecular methods are considered useful and reliable for the identification and differentiation of these species, including conventional PCR, Nested-PCR and Multiplex-PCR. The aim of this study was to optimize PCR techniques for better results in the differentiation of amoeba species from the histolytic complex and Entamoeba hartmanni. For this, 16 positive stool samples were obtained by optical microscopy for Entamoeba histolytica/E. dispar. The results of the present study were similar to those of E. hartmanni, Nested-PCR, and Multiplex-PCR for E. histolytica, E. dispar and E. moshkovskii, with modifications of the original protocols to obtain better results in its standardization. In Conventional PCR two positive samples were observed for E. hartmanni, in the Nested-PCR one sample was positive for E. dispar and for E. moshkovskii species no sample was positive. Although the results of the optimization were not completely satisfactory in all the samples tested, due to the possible degradation of the reagents used, the previous results obtained demonstrate the importance of PCR as an effective tool for the detection and differentiation of species of the genus Entamoeba |
publishDate |
2018 |
dc.date.accessioned.fl_str_mv |
2018-11-07T13:56:18Z |
dc.date.available.fl_str_mv |
2018-11-07T13:56:18Z |
dc.date.issued.fl_str_mv |
2018-10-25 |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/bachelorThesis |
format |
bachelorThesis |
status_str |
publishedVersion |
dc.identifier.citation.fl_str_mv |
Mesquita, Danielle Marcela Menezes de. Otimização da Reação em Cadeia da Polimerase (PCR) para diferenciação molecular de amebas do complexo Histolytica. São Cristóvão, SE, 2018. Monografia (graduação de Biologia) – Departamento de Biologia, Centro de Ciências Biológicas e da Saúde, Universidade Federal de Sergipe, São Cristóvão, 2018 |
dc.identifier.uri.fl_str_mv |
http://ri.ufs.br/jspui/handle/riufs/9595 |
identifier_str_mv |
Mesquita, Danielle Marcela Menezes de. Otimização da Reação em Cadeia da Polimerase (PCR) para diferenciação molecular de amebas do complexo Histolytica. São Cristóvão, SE, 2018. Monografia (graduação de Biologia) – Departamento de Biologia, Centro de Ciências Biológicas e da Saúde, Universidade Federal de Sergipe, São Cristóvão, 2018 |
url |
http://ri.ufs.br/jspui/handle/riufs/9595 |
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por |
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openAccess |
dc.publisher.initials.fl_str_mv |
Universidade Federal de Sergipe |
dc.publisher.department.fl_str_mv |
DBI - Departamento de Biologia – São Cristóvão – Presencial |
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reponame:Repositório Institucional da UFS instname:Universidade Federal de Sergipe (UFS) instacron:UFS |
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