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(Bucharest) Emergence of carbapenemase-producing strains of enterobacteriaceae and their impact on antimicrobial therapy
Romania (FASMR) - University of Medicine and Pharmacy "Carol Davila", Bucharest
Laboratory of Microbiology, National Institute of Infectious Diseases
Prof. Dr. Alexandru Rafila
Dr. Daniela Talapan
Type of Research Project
- Clinical Project with Laboratory work
What is the background of the project?
The infections produced by resistant microorganisms determine a high level of morbidity and mortality due to the therapeutic failure and even bigger costs for medical care. Carbapenem antimicrobials are critically important to human health and they are often the only remaining effective antibiotics for treating serious infections. In the case of Enterobacteriaceae, the resistance to these drugs mediated by acquired carbapenemase enzymes is increasingly encountered in gram-negative bacteria and is rapidly spread because the genes that code it are found on transmissible and mobilisable plasmids. Animal origin food products are recognized as a potential source of resistant organisms (beef, pork and poultry). Considering the fact that raising animals for consumption in Romania is a traditional activity and that pork products are in the top of Romanians’ meat preferences, this research project presents a great importance to our community, mainly aiming to improve the therapeutic protocols in infections with carbapenemase-producing enterobacteriaceae.
What is the aim of the project?
The project aims to find which is the best practice in the microbiology laboratory to identify carbapenemase-producing enterobacteriaceae in order to have access to results rapidly, with affordable prices for testing because the detection of carbapenemase production is complicated (some carbapenemase-producing isolates demonstrate elevated but susceptible, carbapenem Minimum Inhibitory Concentrations- MICs). For the isolation and identification of carbapenemase producers in enterobacteriaceae from different biological specimens, there will mainly be used phenotypic and genotypic methods. Additionally for the comparison of results, specificity and sensitivity tests will also be performed. This should ensure an improvement of the diagnostic capacity and a growth in the accuracy of results. Finally, the research team will verify that it is conducted a correct clinical integration of laboratory data regarding therapeutic protocols in infected patients therapeutic and based on the results of this study, local therapeutic protocols will be enhanced.
What techniques and methods are used?
Gram Staining Technique for the identification of Gram-Negative Bacteria Selective isolation of carbapenem-resistant organisms (Mueller-Hinton Agar plates) Phenotypic and genotypic methods for the identification of carbapenemase-producing strains of enterobacteriaceae • Metallo β-lactamase (MBL) detection (Muller-Hinton agar plate by disk diffusion method) • Modified Hodge’s test (MHT) (Mueller-Hinton agar plate was inoculated with a suspension of a certain strain and streaked for confluent growth using a swab) • PCR analysis Antimicrobial susceptibility testing (Determining Minimum Inhibitory Concentrations by the both micro-dilution and gradient strip methods using E-tests) MAST 4-Disc Tests for the sensitivity and specificity of these types of bacteria (AmpC β-lactamase production)
What is the role of the student?
- The student will observe the practical experiments but will be highly involved in the analysis of the results
What are the tasks expected to be accomplished by the student?
Performing a Gram staining Technique and Interpretating Gram stained smears from cultures Isolation of carbapenem-resistant organisms on Mueller-Hinton Agar plates Testing Antimicrobial susceptibility through disk diffusion, E-test and daily assessment of the inhibition zone PCR technique performed only under supervision and after a clear and detailed explanation of the method PCR analysis along with the research team Clinical integration of the laboratory data
Will there be any theoretical teaching provided (preliminary readings, lectures, courses, seminars etc)
What is expected from the student at the end of the research exchange? What will be the general outcome of the student?
- The student will prepare a scientific report
What skills are required of the student? Is there any special knowledge or a certain level of studies needed?
Basic laboratory skills Motivation Subjects passed: Medical Microbiology, Pharmacology.
Are there any legal limitations in the student’s involvement
Type of students accepted
This project accepts: - Medical students - Graduated students (less than 6 months)
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