Comparação de métodos convencionais e semi-automatizados para identificação de Enterococcus spp.frente à biologia molecular em identificações discrepantes
| Ano de defesa: | 2007 |
|---|---|
| Autor(a) principal: | |
| Orientador(a): | |
| Banca de defesa: | |
| Tipo de documento: | Dissertação |
| Tipo de acesso: | Acesso aberto |
| Idioma: | por |
| Instituição de defesa: |
Não Informado pela instituição
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| Programa de Pós-Graduação: |
Não Informado pela instituição
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| Departamento: |
Não Informado pela instituição
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| País: |
Não Informado pela instituição
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| Palavras-chave em Português: | |
| Link de acesso: | http://www.repositorio.ufc.br/handle/riufc/1764 |
Resumo: | The Enterococcus are Gram-positive cocci catalase negative - which inhabit the gastrointestinal, and genitourinary tract. They are identified manually and the procedure is based on the classic of Facklam, by semi-authomatized and automatized systems as well as by Molecular Biology. Due to the diversity of identification methods and aiming to verify the concordance among some methods, it was accomplished the identification of 62 bacteria strains proceeding from the collection of the Centro Especializado em Micologia Médica (CEMM) of the Federal University of Ceará, in Brazil. They were presumably identified as Enterococcus sp. e Enterococcus faecalis by the Facklam scheme. This study was begun with the identification for two manual methods, modified from the Facklam scheme – Modified 1 and Modified 2- and semi-automatized – API 20 Strep. For the inconsistent results definition we used the semi-automatized – BBL and the Molecular Biology- PRC. The Franklin scheme identified 16% (10) as strains of the genre Enterococcus sp. and 52% (84) as E. faecalis. The Modified Scheme 1 presented the following identification: 82, 2% (51) E. faecalis, 9, 7% (6) E. mundtii and 8, 1% (5) E. gallinarum. The Modified 2 identified 98, 4% (61) E. faecalis and 1, 6% (1) E. gallinarum. The API 20 Strep identified 51,6% (32) E. faecalis, 19,4% (12) A viridans, 13,0% (8) E. avium, 4,8% (3) S. agalactiae, 3,2% (2) E. faecium, 1,6% (1) S. acidominimus, 1,6% (1) Leuconoctocc sp., 1,6% (1) S. uberis, 1,6% (1) A. adiacens and1,6% (1) unacceptable. It was made the selection of 10 sample (strains), among them, the ones numbered (05, 13, 15, 20, 27, 31, 32, 33, 50 and 60), which presented discordant results among the systems Modified 1, Modified 2 and API 20 Strep to be identified by BBL Crystal and by PCR. The BBL identified 6 samples as E. faecium, including the control- strain E. faecalis ATCC 29212 and did not identify 4 samples. In the PCR, one sample did not amplify and 9 were identified as Streptococcus spp. One sample was identified as positive for the genre Enterococcus, but it did not for the species E. faecalis and 1 (one) was amplified for the species S. agalactiae. None of the samples presented were positive for the species E. gallinarum. The control-sample – E. faecalis ATCC 29212 – was correctly amplified. With the analysis of the results, it was noticed that there was identification concordance of the 62 strains in 84% of the samples among the manual systems (Facklam, Modified 1 and Modified 2) and in 52% of the samples among the manual and semi-automatized systems (API 20 Strep). In 10 samples with disagreeing results there was no identification concordance among the manual systems, API 20 Strep and the BBL. A PCR agreed with the manual systems and the BBL and did not agree with the API 20 step, in genre, in one sample. Correlating the PCR with the API 20 Strep, there was agreement, in genre, in 01 sample and disagreement with the other systems. |
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Donato, Silvia TavaresSidrim , José Júlio Costa2012-02-01T14:09:56Z2012-02-01T14:09:56Z2007DONATO, S. T. Comparação de métodos convencionais e semi-automatizados para identificação de Enterococus spp. frente à biologia molecular em identificações discrepantes. 2007. 85 f. Dissertação (Mestrado em Microbiologia Médica)- Universidade Federal do Ceará. Faculdade de Medicina, Fortaleza, 2007.http://www.repositorio.ufc.br/handle/riufc/1764The Enterococcus are Gram-positive cocci catalase negative - which inhabit the gastrointestinal, and genitourinary tract. They are identified manually and the procedure is based on the classic of Facklam, by semi-authomatized and automatized systems as well as by Molecular Biology. Due to the diversity of identification methods and aiming to verify the concordance among some methods, it was accomplished the identification of 62 bacteria strains proceeding from the collection of the Centro Especializado em Micologia Médica (CEMM) of the Federal University of Ceará, in Brazil. They were presumably identified as Enterococcus sp. e Enterococcus faecalis by the Facklam scheme. This study was begun with the identification for two manual methods, modified from the Facklam scheme – Modified 1 and Modified 2- and semi-automatized – API 20 Strep. For the inconsistent results definition we used the semi-automatized – BBL and the Molecular Biology- PRC. The Franklin scheme identified 16% (10) as strains of the genre Enterococcus sp. and 52% (84) as E. faecalis. The Modified Scheme 1 presented the following identification: 82, 2% (51) E. faecalis, 9, 7% (6) E. mundtii and 8, 1% (5) E. gallinarum. The Modified 2 identified 98, 4% (61) E. faecalis and 1, 6% (1) E. gallinarum. The API 20 Strep identified 51,6% (32) E. faecalis, 19,4% (12) A viridans, 13,0% (8) E. avium, 4,8% (3) S. agalactiae, 3,2% (2) E. faecium, 1,6% (1) S. acidominimus, 1,6% (1) Leuconoctocc sp., 1,6% (1) S. uberis, 1,6% (1) A. adiacens and1,6% (1) unacceptable. It was made the selection of 10 sample (strains), among them, the ones numbered (05, 13, 15, 20, 27, 31, 32, 33, 50 and 60), which presented discordant results among the systems Modified 1, Modified 2 and API 20 Strep to be identified by BBL Crystal and by PCR. The BBL identified 6 samples as E. faecium, including the control- strain E. faecalis ATCC 29212 and did not identify 4 samples. In the PCR, one sample did not amplify and 9 were identified as Streptococcus spp. One sample was identified as positive for the genre Enterococcus, but it did not for the species E. faecalis and 1 (one) was amplified for the species S. agalactiae. None of the samples presented were positive for the species E. gallinarum. The control-sample – E. faecalis ATCC 29212 – was correctly amplified. With the analysis of the results, it was noticed that there was identification concordance of the 62 strains in 84% of the samples among the manual systems (Facklam, Modified 1 and Modified 2) and in 52% of the samples among the manual and semi-automatized systems (API 20 Strep). In 10 samples with disagreeing results there was no identification concordance among the manual systems, API 20 Strep and the BBL. A PCR agreed with the manual systems and the BBL and did not agree with the API 20 step, in genre, in one sample. Correlating the PCR with the API 20 Strep, there was agreement, in genre, in 01 sample and disagreement with the other systems.Os Enterococcus são cocos Gram-positivos catalase-negativos, habitantes do trato gastrintestinal e geniturinário. São identificados por meios manuais baseados no clássico de Facklam, por sistemas semi-automatizados, automatizados e por Biologia Molecular. Em vista da diversidade de métodos de identificação e com o intuito de se verificar a concordância entre alguns métodos, foi realizada identificação de 62 cepas bacterianas, advindas da coleção do Centro Especializado em Micologia Médica (CEMM) da Universidade Federal do Ceará, Brasil, presumidamente identificadas como Enterococcus sp. e Enterococcus faecalis pelo esquema de Facklam. Este estudo iniciou-se com a identificação por três métodos manuais: esquema de facklam e dois modificados deste - Modificado 1 e Modificado 2 - e semi-automatizado – API 20 Strep. Para a definição de resultados discordantes, recorreu-se ao semi-automatizado – BBL – e à Biologia Molecular – PCR. O esquema de Facklam identificou 16% (10) como cepas do gênero Enterococcus sp. e 84% (52) como E. faecalis. O esquema Modificado 1 apresentou a seguinte identificação: 82,2% (51) E. faecalis, 9,7% (6) E. mundtii, e 8,1% (5) E. gallinarum. O Modificado 2 identificou 98,4% (61) E. faecalis e 1,6% (1) E. gallinarum. O API 20 Strep identificou 51,6% (32) E. faecalis, 19,4% (12) A viridans, 13,0% (8) E. avium, 4,8% (3) S. agalactiae, 3,2% (2) E. faecium, 1,6% (1) S. acidominimus, 1,6% (1) Leuconoctocc sp., 1,6% (1) S. uberis, 1,6% (1) A. adiacens e 1,6% (1) Inaceitável. Foram selecionadas 10 amostras (cepas n°s 05, 13, 15, 20, 27, 31, 32, 33, 50 e 60), que apresentaram resultados discordantes entre os sistemas Modificado 1, Modificado 2 e API 20 Strep para serem identificadas pelo BBL Crystal e por PCR. O BBL identificou 6 amostras como E. faecium, inclusive a cepa-controle E. faecalis ATCC 29212 e não identificou 4 amostras. Na PCR, uma amostra não amplificou e 9 foram identificadas como Streptococcus spp.. Uma amostra apresentou-se positiva para o gênero Enterococcus, mas não para a espécie E. faecalis e 1 (uma) amplificou para a espécie S. agalactiae. Nenhuma das amostras se apresentou positiva para a espécie E. gallinarum. A amostra-controle – E. faecalis ATCC 29212 - foi amplificada corretamente. Com a análise dos resultados, verificou-se que houve concordância de identificação das 62 cepas em 84% das amostras entre os sistemas manuais (Facklam, Modificado 1 e Modificado 2) e em 52% das amostras entre os sistemas manuais e semi-automatizado (API 20 Strep). Nas 10 amostras com resultados discrepantes, não houve concordância de identificação entre os sistemas manuais, API 20 Strep e o BBL. A PCR concordou com os sistemas manuais e o BBL e foi discordante do API 20 Strep, em gênero, em 01 amostra. Correlacionando a PCR com o API 20 Strep, houve concordância, em gênero, em 01 amostra e foi discordante dos demais sistemas. Para aumentar a sensibilidade de identificação de Enterococcus spp. devem ser utilizados pelo menos dois métodos com testes fenotípicos. Amostras com identificações discordantes devem ser reidentificadas por PCR.Strepto-EnterococcusReação em Cadeia da PolimeraseEnterococcusComparação de métodos convencionais e semi-automatizados para identificação de Enterococcus spp.frente à biologia molecular em identificações discrepantesComparison of convenciaonal and semi-automatized methods to identify Enterococcus spp versus molecular biology in discrepant identificationsinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisporreponame:Repositório Institucional da Universidade Federal do Ceará (UFC)instname:Universidade Federal do Ceará (UFC)instacron:UFCinfo:eu-repo/semantics/openAccessORIGINAL2007_dis_stdonato.pdf2007_dis_stdonato.pdfapplication/pdf1148725http://repositorio.ufc.br/bitstream/riufc/1764/1/2007_dis_stdonato.pdfa3c4b8df96851f37a091b98fb40e19c4MD51LICENSElicense.txtlicense.txttext/plain; charset=utf-81748http://repositorio.ufc.br/bitstream/riufc/1764/2/license.txt8a4605be74aa9ea9d79846c1fba20a33MD52riufc/17642021-02-09 12:04:48.687oai:repositorio.ufc.br:riufc/1764Tk9URTogUExBQ0UgWU9VUiBPV04gTElDRU5TRSBIRVJFClRoaXMgc2FtcGxlIGxpY2Vuc2UgaXMgcHJvdmlkZWQgZm9yIGluZm9ybWF0aW9uYWwgcHVycG9zZXMgb25seS4KCk5PTi1FWENMVVNJVkUgRElTVFJJQlVUSU9OIExJQ0VOU0UKCkJ5IHNpZ25pbmcgYW5kIHN1Ym1pdHRpbmcgdGhpcyBsaWNlbnNlLCB5b3UgKHRoZSBhdXRob3Iocykgb3IgY29weXJpZ2h0Cm93bmVyKSBncmFudHMgdG8gRFNwYWNlIFVuaXZlcnNpdHkgKERTVSkgdGhlIG5vbi1leGNsdXNpdmUgcmlnaHQgdG8gcmVwcm9kdWNlLAp0cmFuc2xhdGUgKGFzIGRlZmluZWQgYmVsb3cpLCBhbmQvb3IgZGlzdHJpYnV0ZSB5b3VyIHN1Ym1pc3Npb24gKGluY2x1ZGluZwp0aGUgYWJzdHJhY3QpIHdvcmxkd2lkZSBpbiBwcmludCBhbmQgZWxlY3Ryb25pYyBmb3JtYXQgYW5kIGluIGFueSBtZWRpdW0sCmluY2x1ZGluZyBidXQgbm90IGxpbWl0ZWQgdG8gYXVkaW8gb3IgdmlkZW8uCgpZb3UgYWdyZWUgdGhhdCBEU1UgbWF5LCB3aXRob3V0IGNoYW5naW5nIHRoZSBjb250ZW50LCB0cmFuc2xhdGUgdGhlCnN1Ym1pc3Npb24gdG8gYW55IG1lZGl1bSBvciBmb3JtYXQgZm9yIHRoZSBwdXJwb3NlIG9mIHByZXNlcnZhdGlvbi4KCllvdSBhbHNvIGFncmVlIHRoYXQgRFNVIG1heSBrZWVwIG1vcmUgdGhhbiBvbmUgY29weSBvZiB0aGlzIHN1Ym1pc3Npb24gZm9yCnB1cnBvc2VzIG9mIHNlY3VyaXR5LCBiYWNrLXVwIGFuZCBwcmVzZXJ2YXRpb24uCgpZb3UgcmVwcmVzZW50IHRoYXQgdGhlIHN1Ym1pc3Npb24gaXMgeW91ciBvcmlnaW5hbCB3b3JrLCBhbmQgdGhhdCB5b3UgaGF2ZQp0aGUgcmlnaHQgdG8gZ3JhbnQgdGhlIHJpZ2h0cyBjb250YWluZWQgaW4gdGhpcyBsaWNlbnNlLiBZb3UgYWxzbyByZXByZXNlbnQKdGhhdCB5b3VyIHN1Ym1pc3Npb24gZG9lcyBub3QsIHRvIHRoZSBiZXN0IG9mIHlvdXIga25vd2xlZGdlLCBpbmZyaW5nZSB1cG9uCmFueW9uZSdzIGNvcHlyaWdodC4KCklmIHRoZSBzdWJtaXNzaW9uIGNvbnRhaW5zIG1hdGVyaWFsIGZvciB3aGljaCB5b3UgZG8gbm90IGhvbGQgY29weXJpZ2h0LAp5b3UgcmVwcmVzZW50IHRoYXQgeW91IGhhdmUgb2J0YWluZWQgdGhlIHVucmVzdHJpY3RlZCBwZXJtaXNzaW9uIG9mIHRoZQpjb3B5cmlnaHQgb3duZXIgdG8gZ3JhbnQgRFNVIHRoZSByaWdodHMgcmVxdWlyZWQgYnkgdGhpcyBsaWNlbnNlLCBhbmQgdGhhdApzdWNoIHRoaXJkLXBhcnR5IG93bmVkIG1hdGVyaWFsIGlzIGNsZWFybHkgaWRlbnRpZmllZCBhbmQgYWNrbm93bGVkZ2VkCndpdGhpbiB0aGUgdGV4dCBvciBjb250ZW50IG9mIHRoZSBzdWJtaXNzaW9uLgoKSUYgVEhFIFNVQk1JU1NJT04gSVMgQkFTRUQgVVBPTiBXT1JLIFRIQVQgSEFTIEJFRU4gU1BPTlNPUkVEIE9SIFNVUFBPUlRFRApCWSBBTiBBR0VOQ1kgT1IgT1JHQU5JWkFUSU9OIE9USEVSIFRIQU4gRFNVLCBZT1UgUkVQUkVTRU5UIFRIQVQgWU9VIEhBVkUKRlVMRklMTEVEIEFOWSBSSUdIVCBPRiBSRVZJRVcgT1IgT1RIRVIgT0JMSUdBVElPTlMgUkVRVUlSRUQgQlkgU1VDSApDT05UUkFDVCBPUiBBR1JFRU1FTlQuCgpEU1Ugd2lsbCBjbGVhcmx5IGlkZW50aWZ5IHlvdXIgbmFtZShzKSBhcyB0aGUgYXV0aG9yKHMpIG9yIG93bmVyKHMpIG9mIHRoZQpzdWJtaXNzaW9uLCBhbmQgd2lsbCBub3QgbWFrZSBhbnkgYWx0ZXJhdGlvbiwgb3RoZXIgdGhhbiBhcyBhbGxvd2VkIGJ5IHRoaXMKbGljZW5zZSwgdG8geW91ciBzdWJtaXNzaW9uLgo=Repositório InstitucionalPUBhttp://www.repositorio.ufc.br/ri-oai/requestbu@ufc.br || repositorio@ufc.bropendoar:2021-02-09T15:04:48Repositório Institucional da Universidade Federal do Ceará (UFC) - Universidade Federal do Ceará (UFC)false |
| dc.title.pt_BR.fl_str_mv |
Comparação de métodos convencionais e semi-automatizados para identificação de Enterococcus spp.frente à biologia molecular em identificações discrepantes |
| dc.title.en.pt_BR.fl_str_mv |
Comparison of convenciaonal and semi-automatized methods to identify Enterococcus spp versus molecular biology in discrepant identifications |
| title |
Comparação de métodos convencionais e semi-automatizados para identificação de Enterococcus spp.frente à biologia molecular em identificações discrepantes |
| spellingShingle |
Comparação de métodos convencionais e semi-automatizados para identificação de Enterococcus spp.frente à biologia molecular em identificações discrepantes Donato, Silvia Tavares Strepto-Enterococcus Reação em Cadeia da Polimerase Enterococcus |
| title_short |
Comparação de métodos convencionais e semi-automatizados para identificação de Enterococcus spp.frente à biologia molecular em identificações discrepantes |
| title_full |
Comparação de métodos convencionais e semi-automatizados para identificação de Enterococcus spp.frente à biologia molecular em identificações discrepantes |
| title_fullStr |
Comparação de métodos convencionais e semi-automatizados para identificação de Enterococcus spp.frente à biologia molecular em identificações discrepantes |
| title_full_unstemmed |
Comparação de métodos convencionais e semi-automatizados para identificação de Enterococcus spp.frente à biologia molecular em identificações discrepantes |
| title_sort |
Comparação de métodos convencionais e semi-automatizados para identificação de Enterococcus spp.frente à biologia molecular em identificações discrepantes |
| author |
Donato, Silvia Tavares |
| author_facet |
Donato, Silvia Tavares |
| author_role |
author |
| dc.contributor.author.fl_str_mv |
Donato, Silvia Tavares |
| dc.contributor.advisor1.fl_str_mv |
Sidrim , José Júlio Costa |
| contributor_str_mv |
Sidrim , José Júlio Costa |
| dc.subject.por.fl_str_mv |
Strepto-Enterococcus Reação em Cadeia da Polimerase Enterococcus |
| topic |
Strepto-Enterococcus Reação em Cadeia da Polimerase Enterococcus |
| description |
The Enterococcus are Gram-positive cocci catalase negative - which inhabit the gastrointestinal, and genitourinary tract. They are identified manually and the procedure is based on the classic of Facklam, by semi-authomatized and automatized systems as well as by Molecular Biology. Due to the diversity of identification methods and aiming to verify the concordance among some methods, it was accomplished the identification of 62 bacteria strains proceeding from the collection of the Centro Especializado em Micologia Médica (CEMM) of the Federal University of Ceará, in Brazil. They were presumably identified as Enterococcus sp. e Enterococcus faecalis by the Facklam scheme. This study was begun with the identification for two manual methods, modified from the Facklam scheme – Modified 1 and Modified 2- and semi-automatized – API 20 Strep. For the inconsistent results definition we used the semi-automatized – BBL and the Molecular Biology- PRC. The Franklin scheme identified 16% (10) as strains of the genre Enterococcus sp. and 52% (84) as E. faecalis. The Modified Scheme 1 presented the following identification: 82, 2% (51) E. faecalis, 9, 7% (6) E. mundtii and 8, 1% (5) E. gallinarum. The Modified 2 identified 98, 4% (61) E. faecalis and 1, 6% (1) E. gallinarum. The API 20 Strep identified 51,6% (32) E. faecalis, 19,4% (12) A viridans, 13,0% (8) E. avium, 4,8% (3) S. agalactiae, 3,2% (2) E. faecium, 1,6% (1) S. acidominimus, 1,6% (1) Leuconoctocc sp., 1,6% (1) S. uberis, 1,6% (1) A. adiacens and1,6% (1) unacceptable. It was made the selection of 10 sample (strains), among them, the ones numbered (05, 13, 15, 20, 27, 31, 32, 33, 50 and 60), which presented discordant results among the systems Modified 1, Modified 2 and API 20 Strep to be identified by BBL Crystal and by PCR. The BBL identified 6 samples as E. faecium, including the control- strain E. faecalis ATCC 29212 and did not identify 4 samples. In the PCR, one sample did not amplify and 9 were identified as Streptococcus spp. One sample was identified as positive for the genre Enterococcus, but it did not for the species E. faecalis and 1 (one) was amplified for the species S. agalactiae. None of the samples presented were positive for the species E. gallinarum. The control-sample – E. faecalis ATCC 29212 – was correctly amplified. With the analysis of the results, it was noticed that there was identification concordance of the 62 strains in 84% of the samples among the manual systems (Facklam, Modified 1 and Modified 2) and in 52% of the samples among the manual and semi-automatized systems (API 20 Strep). In 10 samples with disagreeing results there was no identification concordance among the manual systems, API 20 Strep and the BBL. A PCR agreed with the manual systems and the BBL and did not agree with the API 20 step, in genre, in one sample. Correlating the PCR with the API 20 Strep, there was agreement, in genre, in 01 sample and disagreement with the other systems. |
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2007 |
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2012-02-01T14:09:56Z |
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DONATO, S. T. Comparação de métodos convencionais e semi-automatizados para identificação de Enterococus spp. frente à biologia molecular em identificações discrepantes. 2007. 85 f. Dissertação (Mestrado em Microbiologia Médica)- Universidade Federal do Ceará. Faculdade de Medicina, Fortaleza, 2007. |
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