Random Amplification of Polymorphic DNA Based Typing of Pseudomonas Aeruginosa
P K Menon1, Shiva Priya Eswaran2, S S Pant3
1Classified Specialist (Pathology & Microbiology), Command Hospital (Air Force), Bangalore 560 007.
Abstract:
Pseudomonas aeruginosa was isolated from various sources during the course of an epidemic outbreak of bacterial endophthalmitis following an eye camp at Sangli, Maharashtra. 15 distinct isolates were obtained from clinical samples. Typing of the 15 isolates was performed by random amplified polymorphic DNA (RAPD) analysis, pyocin typing and antibiogram. RAPD typing was rapid, labour friendly and could be done within six hours. RAPD analysis produced reproducible electrophoretic band patterns on the basis of which three distinct amplification patterns could be visualised. The conventional typing methods were labour intensive and took about 48 hours. However, the results of RAPD typing, pyocin typing and antibiogram did not correlate with each other. This study suggests that RAPD typing could be an additional rapid typing method for studying the epidemiology of infectious disease outbreaks due to P aeruginosa.
Insights
This study investigated bacterial endophthalmitis outbreaks caused by Pseudomonas aeruginosa. Random amplified polymorphic DNA (RAPD) typing proved a rapid method for tracking infectious disease outbreaks.
Area of Science:
- Ophthalmology
- Microbiology
- Molecular Biology
Background:
- An epidemic of bacterial endophthalmitis occurred following an eye camp in Sangli, Maharashtra.
- Pseudomonas aeruginosa was identified as the causative agent in clinical samples.
Purpose of the Study:
- To evaluate the utility of Random Amplified Polymorphic DNA (RAPD) analysis as a rapid typing method for Pseudomonas aeruginosa isolates during an epidemic.
- To compare RAPD typing with conventional methods like pyocin typing and antibiogram.
Main Methods:
- Isolation of 15 distinct Pseudomonas aeruginosa strains from clinical samples.
- Typing of isolates using Random Amplified Polymorphic DNA (RAPD) analysis, pyocin typing, and antibiogram.
- RAPD analysis involved reproducible electrophoretic band patterns to identify distinct amplification patterns.
Main Results:
- RAPD typing was significantly faster (within 6 hours) and less labor-intensive than conventional methods (48 hours).
- RAPD analysis revealed three distinct amplification patterns among the isolates.
- There was a lack of correlation between the results obtained from RAPD typing, pyocin typing, and antibiogram.
Conclusions:
- Random Amplified Polymorphic DNA (RAPD) typing is a rapid and effective supplementary method for the epidemiological surveillance of Pseudomonas aeruginosa outbreaks.
- Despite its speed, RAPD typing results did not correlate with conventional typing methods in this study.
- Further research may be needed to understand the discrepancies between different typing methods for Pseudomonas aeruginosa.
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