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Genotyping of Pseudomonas aeruginosa strains isolated from burn patients by RAPD-PCR
Fatemeh Nanvazadeh1, Azar Dokht Khosravi, Mohammad Reza Zolfaghari
1Department of Microbiology, Faculty of Basic Sciences, Qom Branch, Islamic Azad University, Qom, Iran.
Background:
Pseudomonas aeruginosa is one of the important causes of nosocomial infections that easily gains resistance to many antibiotics. This opportunistic pathogen is a major health hazard particularly in immunodeficient patients, patients in intensive care units (ICU) and burn units with life threatening outcome. The bacterium may be originated from different or common sources, and comprises a high colonization and transmission capacity.
Objective:
The aim of present study was to investigate the genotypic variation of Pseudomonas aeroginosa strains isolated from burn patients by using Random Amplified Polymorphic DNA (RAPD) method.
Methods:
Totally 70 clinical samples were collected from burn patients in Taleghani Burn Hospital of Ahvaz. Fifty out of total samples were positive for P. aeruginosa by application of conventional culture and biochemical identification tests. DNA was extracted from the isolates and the RAPD-PCR method was applied to the DNA extracts according to standard method using a short single primer of 272. The technique created repetitive electrophoresis patterns which was used for genotypic differentiation.
Results:
RAPD-PCR, created 9 genotypic profiles designated as I-IX with base pair length ranging from 180 to 2700. Each genotype showed between 3 and 6 different weight DNA bands. Genotype I was the most prevalent, identified in 10 bacterial isolates (20%). Genotypes I, II and VI were mostly common in patients with more severe burn, and were mainly isolated from wound and blood samples obtained from the same patients.
Conclusion:
In present study, we found RAPD-PCR technique as a useful tool for investigation of the genetic variation among P. aeruginosa strains. This is a rapid, low cost, genotypic method with high discriminatory power. The results could assist to screen for the original of infection caused by this organism with subsequent control of colonization and transmission.
Insights
Random Amplified Polymorphic DNA (RAPD)-PCR effectively differentiates Pseudomonas aeruginosa strains from burn patients. This rapid, low-cost method aids in tracking infection sources and controlling transmission of this opportunistic pathogen.
Area of Science:
- Medical Microbiology
- Molecular Biology
- Infectious Diseases
Background:
- Pseudomonas aeruginosa is a significant cause of hospital-acquired infections, particularly in burn patients.
- This opportunistic pathogen exhibits high antibiotic resistance and poses a severe threat to immunocompromised individuals.
- It has a notable capacity for colonization and transmission, often originating from diverse sources.
Purpose of the Study:
- To investigate the genotypic variations among Pseudomonas aeruginosa strains isolated from burn patients.
- To evaluate the utility of the Random Amplified Polymorphic DNA (RAPD) method for strain differentiation.
Main Methods:
- Collected 70 clinical samples from burn patients; 50 were positive for P. aeruginosa.
- Extracted DNA from P. aeruginosa isolates.
- Applied RAPD-PCR using a single primer (272) for genotypic differentiation.
Main Results:
- RAPD-PCR generated 9 distinct genotypic profiles (I-IX) with varying DNA band patterns.
- Genotype I was the most prevalent (20% of isolates).
- Genotypes I, II, and VI were associated with severe burns and frequently isolated from wound and blood samples of the same patients.
Conclusions:
- RAPD-PCR is a valuable tool for assessing genetic diversity in P. aeruginosa strains.
- The technique is rapid, cost-effective, and possesses high discriminatory power.
- Findings can aid in identifying infection origins, thereby controlling colonization and transmission.
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