RAPD- and ERIC-Based Typing of Clinical and Environmental Pseudomonas aeruginosa Isolates
Ibtesam Ghadban Auda1, Israa M S Al-Kadmy1, Sawsan Mohammed Kareem1
1Al-Mustansiriyah University, College of Science, Department of Biology, Branch of Biotechnology, Box 10422, Baghdad, Iraq.
Abstract:
Pseudomonas aeruginosa is a major cause of nosocomial infection in children and adults, resulting in significant morbidity and mortality due to its ability to acquire drug resistance. The ability of P. aeruginosa in the environment to cause infection in individuals has been reported previously; henceforth, surveillance of the emergence and transmission of P. aeruginosa strains among patients is important for infection control in a clinical setup. Various gene-typing methods have been used for epidemiological typing of P. aeruginosa isolates for the purpose of surveillance. In this work, the suitability and comparability of two typing methods, enterobacterial repetitive intergenic consensus (ERIC)-PCR and random amplification of polymorphic DNA (RAPD)-PCR fingerprinting, were studied to characterize P. aeruginosa strains isolated from clinical and environmental sources. Forty-four clinical and environmental bacterial isolates of P. aeruginosa were collected between October 2015 and January 2016. DNA extraction, ERIC-PCR and RAPD-PCR, agarose gel electrophoresis, and phylogenetic analyses were carried using the unweighted pair-group method with mean. RAPD typing revealed less clonality among clinical isolates, whereas the ERIC method showed greater similarity in comparison with RAPD. Environmental isolates, however, showed greater similarity using RAPD compared with ERIC typing. With only a few exceptions, most clinical isolates were distinct from environmental isolates, irrespective of the typing method. In conclusion, both the RAPD and ERIC typing methods proved to be good tools in understanding clonal diversity. The results also suggest that there is no relationship between clinical and environmental isolates. The absence of clonality among the clinical isolates may indicate that most P. aeruginosa infection cases could be endemic and not epidemic and that endemic infections may be due to nonclonal strains of P. aeruginosa.
Insights
This study compared enterobacterial repetitive intergenic consensus (ERIC)-PCR and random amplification of polymorphic DNA (RAPD)-PCR for typing Pseudomonas aeruginosa. Both methods are effective for surveillance, though they revealed different clonal patterns between clinical and environmental isolates.
Area of Science:
- Clinical Microbiology
- Molecular Epidemiology
- Bacterial Genetics
Background:
- Pseudomonas aeruginosa is a significant cause of hospital-acquired infections, leading to severe illness and death, often due to acquired drug resistance.
- Surveillance of P. aeruginosa emergence and transmission is crucial for effective infection control in healthcare settings.
- Gene-typing methods are essential for epidemiological surveillance of P. aeruginosa isolates.
Purpose of the Study:
- To evaluate and compare the suitability of enterobacterial repetitive intergenic consensus (ERIC)-PCR and random amplification of polymorphic DNA (RAPD)-PCR for characterizing P. aeruginosa strains.
- To assess the clonal diversity of P. aeruginosa isolates from both clinical and environmental sources using these two molecular typing methods.
Main Methods:
- Collected 44 clinical and environmental P. aeruginosa isolates.
- Performed DNA extraction, ERIC-PCR, and RAPD-PCR.
- Utilized agarose gel electrophoresis and phylogenetic analyses (unweighted pair-group method with mean) for data interpretation.
Main Results:
- RAPD typing indicated less clonality in clinical isolates compared to ERIC typing.
- ERIC typing showed greater similarity among clinical isolates than RAPD.
- Environmental isolates exhibited higher similarity with RAPD compared to ERIC typing.
- Most clinical isolates were distinct from environmental isolates, regardless of the typing method used.
Conclusions:
- Both RAPD and ERIC typing are valuable tools for understanding the clonal diversity of P. aeruginosa.
- No direct relationship was observed between clinical and environmental P. aeruginosa isolates.
- The lack of clonality in clinical isolates suggests that P. aeruginosa infections are likely endemic rather than epidemic, possibly driven by nonclonal strains.
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