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Artículo
- 1. Felipe Obando Zea Juliana Ocampo Arboleda Third semester– UPB
- 3. INTRODUCTION
- 4. OBJECTIVE -Evaluate genotypic and phenotypic changes of Staphylococcus epidermidis during relapse episodes in prosthetic joint infections.
- 6. METHODS • DNA extraction • PCR
- 7. RESULTS
- 8. RESULTS
- 9. RESULTS
- 10. RESULTS
- 11. DISCUSSIONAuthor Basis Agree or disagree Hischebeth GT, ETAL “On the other hand, isolates were considered as multi-drugresistant and these isolates with this phenotype are considered hard to eradicate, as occurred with methicillin-resistant S. epidermidis in patients with PJI” von Eiff C, ETAL This result is similar to that shown in the patient with pacemaker-associated endocarditis, where S. epidermidis increased the biofilm formation capacity and genetic diversification Dengler Haunreiter V, ETAL it is even assumed that both biofilm formation and SCV phenotype may contribute to the recurrence and persistence of PJI
- 12. CONCLUSION - Diagnosis of Patients - Treatment of Patients - Virus and Bacteria - Antibiotics - Biotechnological Industry
- 14. MIND MAP Staphylococcus Epidermidis specie of bacteria of the Staphylococcaceae family that is part of the normal microbiota of human skin and mucosa along with other species of coagulase-negative staphylococci. Meaning gram-positive, facultative anaerobic and catalase-positive. Microbiology genotypically (pulsed-field gel electrophoresis (PFGE) pattern analysis and genes involved in biofilm formation) phenotypically (antimicrobial resistance, biofilm formation) Bacteria develop resistance mechanisms by using instructions provided by their DNA. Often, resistance genes are found within plasmids, small pieces of DNA that carry genetic instructions from one bacteria to another. This means that some bacteria can share their DNA and make other germs become resistant. technique used for the separation of large deoxyribonucleic acid (DNA) molecules by applying to a gel matrix an electric field that periodically changes direction. Genotypic Antibiotic Resistance PFGE Phenotypic
Editor's Notes
- Staphylococcus epidermidis is a coagulase-negative bacterium capable of causing recurrent relapses in prosthetic joint infection (PJI). The aim of this study was to determine if Staphylococcus epidermidis isolates from patients with recurrent relapses of prosthetic joint infection (PJI) changed genotypically (pulsed-field gel electrophoresis (PFGE) pattern analysis and genes involved in biofilm formation) and phenotypically (antimicrobial resistance, biofilm formation) during the different episodes. Four patients with PJI recurrent relapses were evaluated clinically and microbiologically. Genotypic and phenotypic characteristics of 31 S. epidermidis isolates were determined. In all cases, PJI was treated with antimicrobial therapy and resection of the prosthesis without reimplantation. Months later, all patients had a relapse episode and treated with rifampin plus vancomycin and surgical debridement Prosthetic Joint Infection can be early or late. Early infections is when the infection appears from 0 to 3 months after surgery and they are caused by highly virulent microorganisms like: Staphylococcus aureus and Gramnegative bacteria. Late infections is when the infection occurs from 12 to 24 months after surgery and they are caused by low virulent microorganisms like: Cutibacterium acnes and coagulase-negative staphylococcus. S. epidermidis can be pathogenic, especially when S. epidermidis isolates are biofilm producers and resistant to antibiotics, which are the most common virulence factors.
- Changes in the antibiotics resistance profile in isolates from patients 1 and 2 were observed in the two episodes. Patient 1 had four clones A, B, C, and D that were distributed differentially in the two episodes. Similarly, patients 2 and 3 had two clones and subclones (E-E1 and F-F1, respectively), and patient 4 had only the clone G in both episodes. The clone F formed small-colony variants (SCVs). High level of biofilm formation was found in all clones, except for clones D and G. Clones/ subclones showed a genotypic variation in icaA, sdrF, bap, sesI, and embp genes. The principal coordinate analysis showed that all clones/subclones were different. These results showed that the initial infective clone of S. epidermidis from PJI, changed genotypically and phenotypically after a second relapse as a response to the treatment.
- Antimicrobial susceptibility: Antimicrobial susceptibility tests were carried out in the Vitek 2 computerized system using the sensitivity card AST-GP67. The VITEK system is an automated bacterial identification and antimicrobial susceptibility study. The identification of bacteria is based on the inoculation of a suspension of microorganisms in cards with certain panels of biochemical reactions. Antimicrobial sensitivity is carried out in a similar way through cards containing standardized dilutions of different antibiotics corresponding to the sensitivity cutoff points established. The antibiotics tested were as follows: oxacillin, gentamicin, ciprofloxacin, levofloxacin, erythromycin, clindamycin, linezolid, vancomycin, tetracycline, rifampin, and trimethoprim-sulfamethoxazole. All antimicrobial susceptibility results were analyzed and interpreted with the software Vitek 2 Systems (0.8.01; 2017) using the criteria of Clinical and Laboratory Standards Institute. Only 3 antibiotics were not resistant to the bacteria, there were LZD linezolid, VA vancomycin, RI rifampin. PFGE: or pulsed field gel , samples are made in which electrophoretic profiles are evaluated that give the genotypic expression of a population. In the case of the article, it was used to analyze the genotypic behavior of Staphylococcus epidermidis with the antibiotic.
- DNA Extraction - Basis: use to characterize, isolate and manipulate cellular components, and know its suceptibility or resistance of bacterias to antibiotics. - Why is it use? As a template for the amplification of icaA, ses, embp, and mecA genes, using primers such as: sesA, sesB, sesC, sesD, sesE, sesG, sesH, and embp genes. PCR - Basis: Polimerase chain reaction; process that amplifies (replicates) a small, well-defined segment of DNA many hundreds of thousands of times, creating enough of it for analysis. - Why is it use? ? As a template for the amplification of icaA, ses, embp, and mecA genes, using primers such as: sesA, sesB, sesC, sesD, sesE, sesG, sesH, and embp genes.
- In patient number 1 we can see several clones A, B, C and D. We can see that all the A have the same genetic behavior, however, with clones B, C and D, we can see that they all behave in different ways. In patient number 2 we can see that the E and E1 clones present different genotypic characteristics in some bands. we can observe that the genetic behavior is different because the DNA was modified, this change formed a resistance to antibiotics because it has different genes. that is the reason why the antibiotics should be different in all the clones
- -In Patiente #3 we notice that all Clones F have the same genetic behavior, its DNA does not change and they have an extra gene which helps differentiate them from the other clones. Then we observe that in Clone F1 the genetic behavior is different because the DNA was modified, this means that it formed a resistance to antibiotics because its missing the gene that Clone F has. This means that we cannot order the patient the same antibiotic that was used in the first prosthetic joint infection in the relapsed because it will be a total failure accomplishing its function (killling bacteria) and will be useless. -In Patiente #4 we notice that all Clones G have the same genetic behavior, its DNA is not modified and they have the same quantity of genes. This means that resistance to antiobiotics was not formed, we can treat the patient with the same antibiotic that was used in the first prosthetic joint infection as well in the relapsed period, it would be effective and it will accomplish its function (killing bacteria) successfully.
- Well, in the table we could analyze that the antibiotics that showed the highest resistance to bacteria were oxacyline as we see here in the boxes marked with the red box and gentamicin,
- The genes that have the most resistance to antibiotics in all the patients that were tested are: mecA, sdrG, sesC, sesE, sesF and sesH. They caused what is called a multidrugresistence event that is when each gene gives resistance to a different group of antibiotics. The gene that is less resistant to antibiotics and does not suffers a multidrugresistance event is the sesG, also the gene is really susceptible and sensitive.
- Molecular biology is a great ally of medicine that help us understand the DNA and genetic components to diagnose and treat our patients better. Virus and Bacterias also mutate and its important to know which antibiotic is the best to kill that kind of bacteria, not always the same antibiotic will be effective. In the future molecular biology will keep advancing for the benefit of humankind by helping diagnose new diseases and designing new treatments. The biotechnological industry is now producing hormones, peptides and several vaccines by manipulation of genes in bacteria and cell cultures.
- MAPA CONCEPTUAL JULIANA OCAMPO ARBOLEDA
- FELIPE OBANDO ZEA