Identificación molecular del complejo Burkholderia cepacia, bacteria productora de antibióticos, mediante PCR en tiempo real.
The following research had as the main objective to identify molecularly bacteria producers of antibiotics from the Burkholderia cepacia Complex. They were identified biochemically as Burkholderia cepacia by Egas and Tinajero (2016) with a probability percentage of 88,79% and 99,51% for BC1 and BC2,...
Sábháilte in:
| Príomhchruthaitheoir: | |
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| Formáid: | bachelorThesis |
| Teanga: | spa |
| Foilsithe / Cruthaithe: |
2017
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| Ábhair: | |
| Rochtain ar líne: | http://dspace.ups.edu.ec/handle/123456789/14669 |
| Clibeanna: |
Cuir clib leis
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Cuir clib leis | _version_ | 1840599791847866368 |
|---|---|
| author | Sánchez Robalino, Cristofer Wilson |
| author_facet | Sánchez Robalino, Cristofer Wilson |
| author_role | author |
| collection | Repositorio Universidad Politécnica Salesiana |
| dc.contributor.none.fl_str_mv | Chiluisa Utreras, Viviana Pamela |
| dc.creator.none.fl_str_mv | Sánchez Robalino, Cristofer Wilson |
| dc.date.none.fl_str_mv | 2017-09-28T20:04:27Z 2017-09-28T20:04:27Z 2017-09 |
| dc.format.none.fl_str_mv | application/pdf |
| dc.identifier.none.fl_str_mv | http://dspace.ups.edu.ec/handle/123456789/14669 |
| dc.language.none.fl_str_mv | spa |
| dc.rights.none.fl_str_mv | Atribución-NoComercial-SinDerivadas 3.0 Ecuador http://creativecommons.org/licenses/by-nc-nd/3.0/ec/ info:eu-repo/semantics/openAccess |
| dc.source.none.fl_str_mv | reponame:Repositorio Universidad Politécnica Salesiana instname:Universidad Politécnica Salesiana instacron:UPS |
| dc.subject.none.fl_str_mv | BIOTECNOLOGÍA BURKHOLDERIA CEPACIA BACTERIAS TAXONOMÍA GENOMOVAR |
| dc.title.none.fl_str_mv | Identificación molecular del complejo Burkholderia cepacia, bacteria productora de antibióticos, mediante PCR en tiempo real. |
| dc.type.none.fl_str_mv | info:eu-repo/semantics/publishedVersion info:eu-repo/semantics/bachelorThesis |
| description | The following research had as the main objective to identify molecularly bacteria producers of antibiotics from the Burkholderia cepacia Complex. They were identified biochemically as Burkholderia cepacia by Egas and Tinajero (2016) with a probability percentage of 88,79% and 99,51% for BC1 and BC2, respectively. In order to reach the aim, the methods of extraction and purification of DNA were analyzed statistically by Kit-Roche, Boiling and PCI. The ANOVA test and Tukey determined a significant difference (p<0,001) among the methods implemented. The Kit-Roche methodology presented the best results of quality and concentration of 8,80E+03 and 3,61E+03 (ng/mL) DNA for BC1 and BC2. However, the boiling method is shown as a low cost alternative to demonstrate purity and concentration of 2,40E+03 and 2,41E+03 (ng/mL) DNA for BC1 and BC2, which are acceptable quality characteristics for the Real-time PCR technique. For molecular identification, 16S rRNA-CBc regions, Burkholderia sp. (recA) and B. cepacia-genomovar I (recA) were amplified using specific primers. The analysis of the amplification curves obtained by Real-time PCR confirmed that BC1 belongs to Burkholderia cepacia species (genomovar I), a species that forms the Burkholderia cepacia complex. In turn, it was confirmed that BC2 belongs to the Burkholderia genus, but the genomovar of the organism was not corroborated; however, it is asserted that the strain belongs to some genomovar CBc based on the amplification of the 16S region for CBc and its biochemical identification. |
| eu_rights_str_mv | openAccess |
| format | bachelorThesis |
| id | UPS_75792033c38c3f892249cf816208b51b |
| instacron_str | UPS |
| institution | UPS |
| instname_str | Universidad Politécnica Salesiana |
| language | spa |
| network_acronym_str | UPS |
| network_name_str | Repositorio Universidad Politécnica Salesiana |
| oai_identifier_str | oai:dspace.ups.edu.ec:123456789/14669 |
| publishDate | 2017 |
| reponame_str | Repositorio Universidad Politécnica Salesiana |
| repository.mail.fl_str_mv | . |
| repository.name.fl_str_mv | Repositorio Universidad Politécnica Salesiana - Universidad Politécnica Salesiana |
| repository_id_str | 1737 |
| rights_invalid_str_mv | Atribución-NoComercial-SinDerivadas 3.0 Ecuador http://creativecommons.org/licenses/by-nc-nd/3.0/ec/ |
| spelling | Identificación molecular del complejo Burkholderia cepacia, bacteria productora de antibióticos, mediante PCR en tiempo real.Sánchez Robalino, Cristofer WilsonBIOTECNOLOGÍABURKHOLDERIA CEPACIABACTERIASTAXONOMÍAGENOMOVARThe following research had as the main objective to identify molecularly bacteria producers of antibiotics from the Burkholderia cepacia Complex. They were identified biochemically as Burkholderia cepacia by Egas and Tinajero (2016) with a probability percentage of 88,79% and 99,51% for BC1 and BC2, respectively. In order to reach the aim, the methods of extraction and purification of DNA were analyzed statistically by Kit-Roche, Boiling and PCI. The ANOVA test and Tukey determined a significant difference (p<0,001) among the methods implemented. The Kit-Roche methodology presented the best results of quality and concentration of 8,80E+03 and 3,61E+03 (ng/mL) DNA for BC1 and BC2. However, the boiling method is shown as a low cost alternative to demonstrate purity and concentration of 2,40E+03 and 2,41E+03 (ng/mL) DNA for BC1 and BC2, which are acceptable quality characteristics for the Real-time PCR technique. For molecular identification, 16S rRNA-CBc regions, Burkholderia sp. (recA) and B. cepacia-genomovar I (recA) were amplified using specific primers. The analysis of the amplification curves obtained by Real-time PCR confirmed that BC1 belongs to Burkholderia cepacia species (genomovar I), a species that forms the Burkholderia cepacia complex. In turn, it was confirmed that BC2 belongs to the Burkholderia genus, but the genomovar of the organism was not corroborated; however, it is asserted that the strain belongs to some genomovar CBc based on the amplification of the 16S region for CBc and its biochemical identification.La presente investigación tuvo como objetivo principal identificar molecularmente bacterias productoras de antibióticos del Complejo Burkholderia cepacia; identificadas bioquímicamente como Burkholderia cepacia por Egas y Tinajero (2016), con un porcentaje de probabilidad 88,79% y 99,51% para BC1 y BC2 respectivamente. Para alcanzar el objetivo, se analizaron estadísticamente los métodos de extracción y purificación de ADN: Kit-Roche, Ebullición y PCI, la prueba ANOVA y Tukey determinó una diferencia significativa (p˂0,001) entre los métodos implementados, la metodología Kit-Roche presentó los mejores resultados de calidad y concentración 8,80E+03 y 3,61E+03 (ng/mL) de ADN para BC1 y BC2; sin embargo el método por Ebullición se muestra como una alternativa de bajo costo al presentar pureza y una concentración de 2,40E+03 y 2,41E+03 (ng/mL) de ADN para BC1 y BC2, características de calidad aceptables para la técnica PCR en tiempo real. Para la identificación molecular, se amplificaron las regiones 16S rARN-CBc, Burkholderia sp. (recA) y B. cepacia-genomovar I (recA) empleando primers específicos, el análisis de las curvas de amplificación obtenidas mediante PCR en tiempo real confirmaron que BC1 pertenece a la especie Burkholderia cepacia (genomovar I), especie que conforma el Complejo Burkholderia cepacia; a su vez, se confirmó que BC2 pertenece al: género Burkholderia, pero no se corroboró el genomovar del organismo; sin embargo se asevera que la cepa pertenece algún genomovar del CBc basándonos en la amplificación de la región 16S para CBc y su identificación bioquímica.Chiluisa Utreras, Viviana Pamela2017-09-28T20:04:27Z2017-09-28T20:04:27Z2017-09info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/bachelorThesisapplication/pdfhttp://dspace.ups.edu.ec/handle/123456789/14669spaAtribución-NoComercial-SinDerivadas 3.0 Ecuadorhttp://creativecommons.org/licenses/by-nc-nd/3.0/ec/info:eu-repo/semantics/openAccessreponame:Repositorio Universidad Politécnica Salesianainstname:Universidad Politécnica Salesianainstacron:UPS2019-01-01T19:13:01Zoai:dspace.ups.edu.ec:123456789/14669Institucionalhttps://dspace.ups.edu.ec/Institución privadahttps://www.ups.edu.ec/https://dspace.ups.edu.ec/oai.Ecuador...opendoar:17372019-01-01T19:13:01Repositorio Universidad Politécnica Salesiana - Universidad Politécnica Salesianafalse |
| spellingShingle | Identificación molecular del complejo Burkholderia cepacia, bacteria productora de antibióticos, mediante PCR en tiempo real. Sánchez Robalino, Cristofer Wilson BIOTECNOLOGÍA BURKHOLDERIA CEPACIA BACTERIAS TAXONOMÍA GENOMOVAR |
| status_str | publishedVersion |
| title | Identificación molecular del complejo Burkholderia cepacia, bacteria productora de antibióticos, mediante PCR en tiempo real. |
| title_full | Identificación molecular del complejo Burkholderia cepacia, bacteria productora de antibióticos, mediante PCR en tiempo real. |
| title_fullStr | Identificación molecular del complejo Burkholderia cepacia, bacteria productora de antibióticos, mediante PCR en tiempo real. |
| title_full_unstemmed | Identificación molecular del complejo Burkholderia cepacia, bacteria productora de antibióticos, mediante PCR en tiempo real. |
| title_short | Identificación molecular del complejo Burkholderia cepacia, bacteria productora de antibióticos, mediante PCR en tiempo real. |
| title_sort | Identificación molecular del complejo Burkholderia cepacia, bacteria productora de antibióticos, mediante PCR en tiempo real. |
| topic | BIOTECNOLOGÍA BURKHOLDERIA CEPACIA BACTERIAS TAXONOMÍA GENOMOVAR |
| url | http://dspace.ups.edu.ec/handle/123456789/14669 |