Document Type : Research Paper
Authors
Tikrit university - College of science.
Abstract
The aim of the present study was the molecular characterization and the evaluation of variability and genetic relationship of six Pseudomonas aeruginosa isolates using PCR-based Randomly Amplified Polymorphic DNA (RAPD) technique. A total number of 86 samples were collected from patients that hospitalized in Tikrit Teaching Hospital in Tikrit city. These samples were taken from patients basing on the sources of infections, the isolates were taken from: wounds, ear, burns, urine, sputum, and eyes infections. Using enrichment, selective media, and biochemical tests, that characterized and identified as P. aeruginosa.
Genomic DNA was extracted from six P. aeruginosa isolates isolated from these different sources. These genomic DNA samples were found to have a suitable concentration and purity for RAPD-PCR analysis. RAPD-PCR technique was performed using 15 different Operon random primers. Eleven primers gave successful amplification results in repeated experiments. As a result, the total number of amplified bands was 270 and the total number of polymorphic bands was 234. The highest number of polymorphic bands (39 bands) was produced by primer OPX-01. The primer efficiency ranged from 3.70 (primer OPA-11) to 14.44 (primer OPX-01) and the discriminatory value ranged from 1.70% (primer OPA-11) to 16.66% (primer OPX-01). In addition, genetic distance and cluster analysis among different P. aeruginosa isolates were estimated by using UPGMA computer program basing on RAPD-PCR banding patterns that obtained in this study. These results suggesting that possible and frequent occurrence of mutations in DNA sequencing P. aeruginosa bacteria from different sources and locations. This study has proved existence genetic differences (DNA polymorphism) among the six P. aeruginosa isolates isolated from different sources. Therefore, we can say that RAPD technique could be an efficient technique for studying the molecular characterization and the epidemiology of P. aeruginosa bacteria.
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