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 B...
| Autor Principal: | Sánchez Robalino, Cristofer Wilson |
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| Formato: | bachelorThesis |
| Idioma: | spa |
| Publicado: |
2017
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| Materias: | |
| Acceso en línea: |
http://dspace.ups.edu.ec/handle/123456789/14669 |
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| Sumario: |
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. |
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