Diversity among clinical isolates of Proteus penneri detected by random amplified polymorphic DNA analysis
G Hoffmann1, G Gajdos, M Czakó
1Department of Genetics and Microbiology, Janus Pannonius University, Pécs, Hungary.
Abstract:
DNA of thirteen haemolytic Proteus penneri strains of clinical origin, all producing calcium dependent haemolysin and having been derived from four European countries was examined for plasmid profile, and outer membrane protein profile, by random amplified polymorphic DNA-PCR (RAPD-PCR) method, and digestions with restriction endonucleases were performed. All strains contained two large plasmids of approximately 60 and 70 kilobase pairs (kb). In addition, four strains contained a small plasmid of about 6 kb. These four strains produced cell-bound haemolysin only. Outer membrane protein analysis revealed subtle differences between strains. RAPD-PCR with primer I (CCGCAGCCAA) revealed 13 types, whereas primer II (AACGCGCAAC) yielded only two main types of different patterns. Results with primer I suggests a DNA sequence diversity within this species. The RAPD-PCR method provides a fast, economical and reproducible means for the typing of P. penneri. Digestion with restriction endonucleases indicated a high level of DNA methylation in this species.
Insights
This study analyzed the DNA of thirteen Proteus penneri strains, revealing genetic diversity through plasmid profiles and outer membrane proteins. Random amplified polymorphic DNA-PCR (RAPD-PCR) proved effective for strain typing.
Area of Science:
- Microbiology
- Molecular Biology
- Genetics
Background:
- Haemolytic Proteus penneri strains of clinical origin are significant pathogens.
- Calcium-dependent haemolysin production is a key virulence factor in these strains.
- Understanding the genetic relatedness and diversity of Proteus penneri is crucial for epidemiological studies.
Purpose of the Study:
- To investigate the genetic diversity and relatedness of thirteen haemolytic Proteus penneri strains from clinical origins.
- To evaluate the utility of Random Amplified Polymorphic DNA-PCR (RAPD-PCR) for typing Proteus penneri.
- To characterize plasmid profiles and outer membrane proteins of these strains.
Main Methods:
- DNA analysis of thirteen Proteus penneri strains.
- Plasmid profiling and outer membrane protein analysis.
- Random Amplified Polymorphic DNA-PCR (RAPD-PCR) using two different primers.
- Restriction endonuclease digestion analysis.
Main Results:
- All strains possessed two large plasmids (approx. 60 and 70 kb); four strains also had a small plasmid (approx. 6 kb).
- Outer membrane protein analysis showed subtle inter-strain variations.
- RAPD-PCR with primer I identified 13 distinct strain types, indicating significant DNA sequence diversity.
- Primer II yielded only two main patterns, while restriction endonuclease digestion suggested high DNA methylation.
Conclusions:
- RAPD-PCR is a rapid, cost-effective, and reproducible method for typing Proteus penneri strains.
- The study highlights genetic diversity within Proteus penneri species.
- Plasmid profiles and protein analysis provide additional discriminatory markers for strain characterization.
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