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SEMINARIO BIOLOGÍA MOLECULAR
- 1. MARIA CLARA VARGAS ZULUAGA THIRD SEMESTER UPB
- 2. About Acinetobacter baumannii INTRODUCTION • It´s an aerobic Gram-negative bacterium. • It is estimated that up to 10% of nosocomial infections in the USA and 2% in Europe are caused by this pathogen, with these frequencies almost doubling in Asia and the Middle East. • 45% of A. baumanii exhibit multidrug resistance (MDR) globally. • It´s considered a priority pathogen by WHO. https://es.wikipedia.org/wiki/Acinetobacter_baumannii
- 3. INTRODUCTION Multidrug Resistance (MDR) A microorganism it´s considered multiresistant when it´s shows resistance to 3 different categories of antimicrobial drugs. They do it via spontaneous mutation or DNA transfer. • Cell envelope as a barrier • Multidrug efflux systems • Mutations in genes coding for porins and antibiotic targets • Enzymes that degrade/inactivate antibiotics.
- 4. OBJECTIVE To develop a scar-free genome-editing tool suitable for chromosomal and plasmid modifications in MDR A. baumannii AB5075
- 5. METHODS Triparental mating - PCR Plasmid construction https://es.khanacademy.org/science/ap-biology/gene-expression-and- regulation/biotechnology/a/overview-dna-cloning The gene is inserted into a circular piece of DNA (plasmid). Insertion is done with enzymes, resulting in a recombinant DNA molecule, which is DNA assembled from fragments from multiple sources. PCR involves using short pieces of synthetic DNA, called primers, to select a segment of the genome to be amplified, and then multiple DNA synthesis sessions to amplify that segment. https://www.genome.gov/es/genetics-glossary/Reaccion-en-cadena-de-la-polimerasa
- 6. METHODS Antibiotic disc diffusion assay (DDA) MIC determination Used to determine the susceptibility of clinical isolates of bacteria to different antibiotics Is the lowest concentration of an antimicrobial that will inhibit the visible growth of a microorganism after overnight incubation
- 7. RESULTS
- 8. RESULTS
- 9. Vila J, Martí S, Sánchez-Céspedes J. Porins Whereas there are reports about the role of CraA in chloramphenicol resistance, the RI2-encoded CmlA5 has only been annotated as a chloramphenicol efflux pump based on homology Agree Anderson SE, Sherman EX, Weiss DS Homologous recombination between the two integrase regions has been documented before, amplifying the copy number of RI2 and producing aminoglycoside heteroresistance. Agree Foong WE, Tam HK, Crames JJ Apart from its role in chloramphenicol resistance,35A. baumannii CraA was recently shown to have a broader substrate range, including other chloramphenicol derivates and biocides, such as chlorhexidine, benzalkonium and dequalinium Agree DISCUSSION
- 10. CONCLUSSIONS - Molecular biology is important for the study of microorganisms that can be a threat to the hospital environment and ultimately the treatment of patients. - It is important that studies continue with molecular biology methods to understand antimicrobial resistance mechanisms and how to overcome them to avoid possible incurable mutations.
Editor's Notes
- Explicar q es gran negativo Nosocomial: Nosocomial infections also referred to as healthcare-associated infections (HAI), are infection(s) acquired during the process of receiving health care that was not present during the time of admission MDR: resistance to at least three classes of antibiotics), DECIR SOBRE MDR: These include the cell envelope as a barrier, multidrug efflux systems and mutations in genes coding for porins and antibiotic targets (e.g. ribosomal proteins, PBPs, DNA replication enzymes and the lipid A biosynthetic pathway), as well as enzymes that degrade/inactivate antibiotics.
- These include the cell envelope as a barrier, multidrug efflux systems and mutations in genes coding for porins and antibiotic targets (e.g. ribosomal proteins, PBPs, DNA replication enzymes and the lipid A biosynthetic pathway), as well as enzymes that degrade/inactivate antibiotics.