Related Experiment Videos
[Molecular cloning of the imipenem resistant gene in bacteroides fragilis]
Abstract:
Imipenem, as a representative of-cabapenem, is one of the most effective beta-Lactam agents against Bacteroides, but recently there have been a few reports on the resistant strains of Bacteroides. To explore the mechanism of imipenem resistance (IMPr) at the molecular level, we extracted chromosome DNA of the resistant strains and acquired a 1.7 kb fragment of resistant gene of imipenem. Recombinant plasmid was stably reserved in generation after generation in E. coli DH5. Resistant gene of imipenem was labeled with Dig-11-dUTP by way of random prime, and 40 strains of clinically isolated Bacteroides were detected by the prepared DNA probe. Two IMPr strains showed hybridization signal, whereas 38 IMPr strains did not show such a signal. The result suggests that the Dig-labeled IMPr gene probe can be used in molecular epidemiology for investigating IMPr strains.
Insights
A novel DNA probe successfully identified imipenem resistance (IMPr) genes in Bacteroides strains. This molecular tool aids in tracking imipenem-resistant Bacteroides in clinical settings.
Area of Science:
- Microbiology
- Molecular Biology
- Genetics
Context:
- Carbapenem antibiotics, like imipenem, are crucial for treating infections caused by Bacteroides species.
- Emerging resistance to imipenem in Bacteroides poses a significant clinical challenge.
- Understanding the molecular basis of imipenem resistance (IMPr) is essential for effective treatment strategies.
Purpose:
- To explore the molecular mechanisms underlying imipenem resistance in Bacteroides.
- To develop and validate a DNA probe for detecting imipenem resistance genes.
- To assess the utility of the developed probe in molecular epidemiology.
Summary:
- Researchers isolated and characterized a 1.7 kb DNA fragment conferring imipenem resistance from resistant Bacteroides strains.
- This fragment was used to create a Dig-labeled DNA probe.
- The probe successfully identified imipenem resistance genes in 2 out of 40 clinically isolated Bacteroides strains, suggesting a specific genetic basis for resistance in these cases.
Impact:
- The developed Dig-labeled DNA probe demonstrates potential for molecular epidemiological surveillance of imipenem-resistant Bacteroides.
- This tool can aid in rapid identification and characterization of resistant strains.
- Facilitates targeted interventions to control the spread of antibiotic resistance.