Abstract
Molecular techniques, being fast and precise, have been used in the taxonomic research of many bacterial genera. The identification of a very heterogeneous group of species, such as the Burkholderia cepacia complex (Bcc) and related genera, through basic microbiology techniques, have been insufficient. In recent years, however, several studies have identified the biotechnological potential of (Bcc). Thus, generating techniques such as real-time PCR to be used as a definitive typing tool has been of utmost importance in research. The main objective of the present investigation was to identify bacteria producing (Bcc) antibiotics molecularly, through the analysis of the RecA region by means of real-time PCR. To fulfil with the objective, DNA extraction and purification methods were analyzed statistically: High Pure Template Preparation Kit-ROCHE, boiling and PCI, by means of ANOVA and Tukey, determining a significant difference (p˂0.001); The ROCHE kit methodology presented the best DNA quality and concentration results. The boiling method is shown here, as a low-costalternative to show DNA with acceptable characteristics and quality for the real-time PCR technique. For molecular identification, amplification of the 16S regions rRNA-Bcc, Burkholderia sp. (RecA) and B. cepacia-genomovar I (RecA) using specific primers took place. The analysis of the amplification curves obtained by real-time PCR confirmed that BC1 belongs to the species Burkholderia cepacia (genomovar I); afterwards, it was confirmed that BC2 belongs to the genus Burkholderia, but the genomovar of the organism was not corroborated; however it is asserted that the strain belongs to some genomovar of Bcc based on the amplification of the 16S region for Bcc and its biochemical identification.
Translated title of the contribution | Identificación Molecular de la Bacteria Productora de Antibióticos, Complejo Burkholderia Cepacia, Mediante Pcr en Tiempo Real |
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Original language | English (US) |
Pages (from-to) | 97-103 |
Number of pages | 7 |
Journal | Internacional Journal Of Bioprocess & Biotectonological Advancements |
Volume | 3 |
Issue number | 3 |
State | Published - 2 Jan 2018 |
Keywords
- Burkholderia cepacia complex
- Genomovar
- Molecular detection
- Real-time pcr
- Reca gene
CACES Knowledge Areas
- 115A Biology