Reação em cadeia da polimerase no diagnóstico e prevalência do papilomavírus

Detalhes bibliográficos
Autor(a) principal: Ribeiro Júnior, José Pires
Data de Publicação: 2001
Tipo de documento: Dissertação
Idioma: por
Título da fonte: Repositório Institucional da UFU
Texto Completo: https://repositorio.ufu.br/handle/123456789/29710
http://doi.org/10.14393/ufu.di.2001.45
Resumo: Câncer of the uterine cervix is the second cause of death among women worldwide. Among the risk factors for cervical câncer, human papillomavirus (HPV) is the most important one, being present in approximately 100% of the tumors. The polymerase chain reaction (PCR) is the most sensitive test for the diagnosis of HPV in the female genital tract, but factors such as primers and detection systens may affect PCR results, and the final diagnosis of HPV infection. OBJECTIVES: To evaluate the sensitivity of agarose and polyacrylamide gels in the diagnosis of HPV by PCR in smears from the human uterine cervix, to determine the prevalence of HPV by combining the two pairs of primers (MY09/MY11 and GP5+/GP6+), and to compare the results with those obtained with each primer separately. MATERIAL AND METHOD: Eighty-six samples of cervix secretion were studied. Mean patient age was 32 years and 6 months. Each sample was submitted to four PCR procedures using the specific primers MY09/MY11 and GP5+/GP6+ which amplify conserved L1 regions. and then evaluated on 1.5% agarose gel ( stained with Ethidium bromide and visualized under the UV light) and later visualized on 10% polyacrylamide gel (stained with silver nitrate). Data were analyzed statistically by the McMenar test, the z test and the Kappa index, with the levei of significance of p = 0.05.XIV RESULTS: Of the samples from the 86 patients evaluated, 14 (16.27%) were positive to MY09/MY11 on the agarose gel and 20 (23.25%) were positive on the polyacrylamide gel, which detected all the samples that were positive on the agarose gel plus six new ones. With the GP5+/GP6+ primer, 15 samples (17.44%) were positive on agarose gel and 38 (44.18%) were positive on polyacrylamide gel, which detected all the samples that were positive on the agarose gel plus 23 new ones. When the two primers were used in combination, the prevalence was 31.39% on agarose gel with 27 positive samples, but the total prevalence was 48 positive samples (55.81%), obtained with the combination of the two pairs of primers on polyacrylamide gel. CONCLUSION: The polyacrylamide gel presented greater resolution on than the agarose gel system with both sets of primers, GP+ and MY, for the HPV detection. The combination of both markers and gel systens increased HPV prevalence.
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spelling Reação em cadeia da polimerase no diagnóstico e prevalência do papilomavírusPolymerase chain reaction in diagnosis and prevalence papillomavirusCâncer de colo uterinoPapilomavírus humanoCNPQ::CIENCIAS DA SAUDEPapilomavírusReação em cadeia de polimeraseDiagnósticoColo uterino - CâncerCâncer of the uterine cervix is the second cause of death among women worldwide. Among the risk factors for cervical câncer, human papillomavirus (HPV) is the most important one, being present in approximately 100% of the tumors. The polymerase chain reaction (PCR) is the most sensitive test for the diagnosis of HPV in the female genital tract, but factors such as primers and detection systens may affect PCR results, and the final diagnosis of HPV infection. OBJECTIVES: To evaluate the sensitivity of agarose and polyacrylamide gels in the diagnosis of HPV by PCR in smears from the human uterine cervix, to determine the prevalence of HPV by combining the two pairs of primers (MY09/MY11 and GP5+/GP6+), and to compare the results with those obtained with each primer separately. MATERIAL AND METHOD: Eighty-six samples of cervix secretion were studied. Mean patient age was 32 years and 6 months. Each sample was submitted to four PCR procedures using the specific primers MY09/MY11 and GP5+/GP6+ which amplify conserved L1 regions. and then evaluated on 1.5% agarose gel ( stained with Ethidium bromide and visualized under the UV light) and later visualized on 10% polyacrylamide gel (stained with silver nitrate). Data were analyzed statistically by the McMenar test, the z test and the Kappa index, with the levei of significance of p = 0.05.XIV RESULTS: Of the samples from the 86 patients evaluated, 14 (16.27%) were positive to MY09/MY11 on the agarose gel and 20 (23.25%) were positive on the polyacrylamide gel, which detected all the samples that were positive on the agarose gel plus six new ones. With the GP5+/GP6+ primer, 15 samples (17.44%) were positive on agarose gel and 38 (44.18%) were positive on polyacrylamide gel, which detected all the samples that were positive on the agarose gel plus 23 new ones. When the two primers were used in combination, the prevalence was 31.39% on agarose gel with 27 positive samples, but the total prevalence was 48 positive samples (55.81%), obtained with the combination of the two pairs of primers on polyacrylamide gel. CONCLUSION: The polyacrylamide gel presented greater resolution on than the agarose gel system with both sets of primers, GP+ and MY, for the HPV detection. The combination of both markers and gel systens increased HPV prevalence.Dissertação (Mestrado)Em todo o mundo o câncer de colo uterino humano e a segunda causa de morte entre as mulheres. Entre os fatores de risco para o câncer cervical o Papilomavírus Humano (HPV) é o mais importante, estando presente em aproximadamente 100% dos tumores. A Reação em Cadeia da Polimerase (PCR) é o exame mais sensível no diagnóstico do HPV no trato genital feminino, porém análise de fatores como primers e géis de leitura utilizado na PCR são escassos e podem, influenciar no diagnóstico final da infeção pelo HPV. OBJETIVOS: Avaliar a sensibilidade dos géis de leitura de Agarose e de Poliacrilamida no diagnóstico de HPV pela Reação em Cadeia da Polimerase em escovado de colo uterino humano, bem como determinar a prevalência do HPV associando os dois pares de primers (MY09/MY11 e GP5+/GP6+) e comparar com os resultados de cada primer isoladamente MATERIAL E MÉTODO: Foram estudadas 86 amostras de secreção do colo uterino. A media das idades das pacientes foram de 32 anos e 6 meses. Cada amostra foi submetida a quatro reações de PCR, usando primers específicos: MY09/MY11 e GP5+/GP6+ que amplificam regiões L1 conservadas e, em seguida, foram avaliados em gel de agarose a 1,5% e posteriormente visualizados em gel de poliacrilamida a 10% (corados com nitrato de prata). Para a análise estatística foi aplicada a prova de McMenar, o teste z e o índice de Kappa com nível de significância de p= 0,05.Xll RESULTADOS: Das 86 pacientes avaliadas, 14 amostras (16,27%) foram positivas em MY09/MY11 no gel de agarose e 20 (23,25%) foram positivas no gel de poliacriiamida, sendo que este gel detectou todas as amostras positivas no gel de agarose e mais seis novas. Com o primer GP5+/GP6+ 15 amostras (17,44%) foram positivas em gel de agarose e 38 amostras (44,18%) foram positivas em gel de poliacriiamida, sendo que este detectou todas as amostras do gel de agarose e mais 23 novas amostras. A prevalência usando a associação dos dois prímers em gel de agarose foi de 31,39% com 27 amostras positivas, porém a prevalência total foi de 48 amostras positivas (55,81%), sendo este dado obtido com associação dos dois pares de prímers em gel de poliacriiamida. CONCLUSÕES: O gel de poliacriiamida foi melhor que o gel de agarose tanto no primer GP+-PCR, quanto no primer MY-PCR para diagnóstico do HPV. A associação dos prímers GP+-PCR e MY-PCR aumentou a positividade do diagnóstico do HPV pela PCR nos dois géis de leitura utilizado.Universidade Federal de UberlândiaBrasilPrograma de Pós-graduação em Ciências da SaúdeAchá, Renato Enrique Sologurenhttp://lattes.cnpq.br/8612786219015145Goulart Filho, Luiz Ricardohttp://lattes.cnpq.br/6759395798493082Ribeiro Júnior, José Pires2020-08-18T11:47:15Z2020-08-18T11:47:15Z2001info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisapplication/pdfRIBEIRO JÚNIOR, José Pires. Reação em cadeia da polimerase no diagnóstico e prevalência do papilomavírus. 2001.78 f. Dissertação (Mestrado em Ciências da Saúde) - Universidade Federal de Uberlândia, Uberlândia, 2020. DOI http://doi.org/10.14393/ufu.di.2001.45https://repositorio.ufu.br/handle/123456789/29710http://doi.org/10.14393/ufu.di.2001.45porhttp://creativecommons.org/licenses/by-nc-nd/3.0/us/info:eu-repo/semantics/openAccessreponame:Repositório Institucional da UFUinstname:Universidade Federal de Uberlândia (UFU)instacron:UFU2020-08-19T06:19:49Zoai:repositorio.ufu.br:123456789/29710Repositório InstitucionalONGhttp://repositorio.ufu.br/oai/requestdiinf@dirbi.ufu.bropendoar:2020-08-19T06:19:49Repositório Institucional da UFU - Universidade Federal de Uberlândia (UFU)false
dc.title.none.fl_str_mv Reação em cadeia da polimerase no diagnóstico e prevalência do papilomavírus
Polymerase chain reaction in diagnosis and prevalence papillomavirus
title Reação em cadeia da polimerase no diagnóstico e prevalência do papilomavírus
spellingShingle Reação em cadeia da polimerase no diagnóstico e prevalência do papilomavírus
Ribeiro Júnior, José Pires
Câncer de colo uterino
Papilomavírus humano
CNPQ::CIENCIAS DA SAUDE
Papilomavírus
Reação em cadeia de polimerase
Diagnóstico
Colo uterino - Câncer
title_short Reação em cadeia da polimerase no diagnóstico e prevalência do papilomavírus
title_full Reação em cadeia da polimerase no diagnóstico e prevalência do papilomavírus
title_fullStr Reação em cadeia da polimerase no diagnóstico e prevalência do papilomavírus
title_full_unstemmed Reação em cadeia da polimerase no diagnóstico e prevalência do papilomavírus
title_sort Reação em cadeia da polimerase no diagnóstico e prevalência do papilomavírus
author Ribeiro Júnior, José Pires
author_facet Ribeiro Júnior, José Pires
author_role author
dc.contributor.none.fl_str_mv Achá, Renato Enrique Sologuren
http://lattes.cnpq.br/8612786219015145
Goulart Filho, Luiz Ricardo
http://lattes.cnpq.br/6759395798493082
dc.contributor.author.fl_str_mv Ribeiro Júnior, José Pires
dc.subject.por.fl_str_mv Câncer de colo uterino
Papilomavírus humano
CNPQ::CIENCIAS DA SAUDE
Papilomavírus
Reação em cadeia de polimerase
Diagnóstico
Colo uterino - Câncer
topic Câncer de colo uterino
Papilomavírus humano
CNPQ::CIENCIAS DA SAUDE
Papilomavírus
Reação em cadeia de polimerase
Diagnóstico
Colo uterino - Câncer
description Câncer of the uterine cervix is the second cause of death among women worldwide. Among the risk factors for cervical câncer, human papillomavirus (HPV) is the most important one, being present in approximately 100% of the tumors. The polymerase chain reaction (PCR) is the most sensitive test for the diagnosis of HPV in the female genital tract, but factors such as primers and detection systens may affect PCR results, and the final diagnosis of HPV infection. OBJECTIVES: To evaluate the sensitivity of agarose and polyacrylamide gels in the diagnosis of HPV by PCR in smears from the human uterine cervix, to determine the prevalence of HPV by combining the two pairs of primers (MY09/MY11 and GP5+/GP6+), and to compare the results with those obtained with each primer separately. MATERIAL AND METHOD: Eighty-six samples of cervix secretion were studied. Mean patient age was 32 years and 6 months. Each sample was submitted to four PCR procedures using the specific primers MY09/MY11 and GP5+/GP6+ which amplify conserved L1 regions. and then evaluated on 1.5% agarose gel ( stained with Ethidium bromide and visualized under the UV light) and later visualized on 10% polyacrylamide gel (stained with silver nitrate). Data were analyzed statistically by the McMenar test, the z test and the Kappa index, with the levei of significance of p = 0.05.XIV RESULTS: Of the samples from the 86 patients evaluated, 14 (16.27%) were positive to MY09/MY11 on the agarose gel and 20 (23.25%) were positive on the polyacrylamide gel, which detected all the samples that were positive on the agarose gel plus six new ones. With the GP5+/GP6+ primer, 15 samples (17.44%) were positive on agarose gel and 38 (44.18%) were positive on polyacrylamide gel, which detected all the samples that were positive on the agarose gel plus 23 new ones. When the two primers were used in combination, the prevalence was 31.39% on agarose gel with 27 positive samples, but the total prevalence was 48 positive samples (55.81%), obtained with the combination of the two pairs of primers on polyacrylamide gel. CONCLUSION: The polyacrylamide gel presented greater resolution on than the agarose gel system with both sets of primers, GP+ and MY, for the HPV detection. The combination of both markers and gel systens increased HPV prevalence.
publishDate 2001
dc.date.none.fl_str_mv 2001
2020-08-18T11:47:15Z
2020-08-18T11:47:15Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/masterThesis
format masterThesis
status_str publishedVersion
dc.identifier.uri.fl_str_mv RIBEIRO JÚNIOR, José Pires. Reação em cadeia da polimerase no diagnóstico e prevalência do papilomavírus. 2001.78 f. Dissertação (Mestrado em Ciências da Saúde) - Universidade Federal de Uberlândia, Uberlândia, 2020. DOI http://doi.org/10.14393/ufu.di.2001.45
https://repositorio.ufu.br/handle/123456789/29710
http://doi.org/10.14393/ufu.di.2001.45
identifier_str_mv RIBEIRO JÚNIOR, José Pires. Reação em cadeia da polimerase no diagnóstico e prevalência do papilomavírus. 2001.78 f. Dissertação (Mestrado em Ciências da Saúde) - Universidade Federal de Uberlândia, Uberlândia, 2020. DOI http://doi.org/10.14393/ufu.di.2001.45
url https://repositorio.ufu.br/handle/123456789/29710
http://doi.org/10.14393/ufu.di.2001.45
dc.language.iso.fl_str_mv por
language por
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info:eu-repo/semantics/openAccess
rights_invalid_str_mv http://creativecommons.org/licenses/by-nc-nd/3.0/us/
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Universidade Federal de Uberlândia
Brasil
Programa de Pós-graduação em Ciências da Saúde
publisher.none.fl_str_mv Universidade Federal de Uberlândia
Brasil
Programa de Pós-graduação em Ciências da Saúde
dc.source.none.fl_str_mv reponame:Repositório Institucional da UFU
instname:Universidade Federal de Uberlândia (UFU)
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instname_str Universidade Federal de Uberlândia (UFU)
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reponame_str Repositório Institucional da UFU
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repository.name.fl_str_mv Repositório Institucional da UFU - Universidade Federal de Uberlândia (UFU)
repository.mail.fl_str_mv diinf@dirbi.ufu.br
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