Rapid High Specific Method for the Detection of Pseudomonas fluorescens

Mohammed Abdullatif Hamod^1,

¹Department of Biotechnology, College of Science, University of Baghdad, Baghdad, Iraq

Cite this paper:
Mohammed Abdullatif Hamod. Rapid High Specific Method for the Detection of Pseudomonas fluorescens. American Journal of Microbiological Research. 2015; 3(5):160-164. doi: 10.12691/ajmr-3-5-2

Abstract

The molecular detection Pseudomonas fluorescens has a vital importance for clinical and environmental microbiologist. Therefore this work aimed to establish a specific and sensitive method for diagnosis of this bacterium. To perform this aim ten Ps. fluorescens, eight closely related Pseudomonas isolates and another four common bacteria (Escherichia coli, Proteus mirabilis, klebsiella pneumonia and Bacillus sp) were collected and their detection were confirmed by 16S rRNA sequencing. Scanning the complete genome of Ps. fluorescens available in GenBank have denoted for the distinction of cumene dioxygenase. A couple of primer was designed to amplify 498 bp of cumene dioxygenase from Ps. fluorescens. The PCR using these primers resulted in single band when DNA purified from Ps. fluorescens isolates were used as template more over none of the of the DNA extracted from other isolates used in the study was resulting in any band at the experiment condition. The targeted band was amplified when serial dilution of Ps. fluorescens DNA (400, 200, 100, 50, 25, 12, 6, 3, 2, 1, 0.5 ng /μl) used as template in PCR pointing the high sensitivity of the method. Therefore this work is presenting a sensitive and specific method for the detection of Ps. fluorescens.

Keywords:
Ps. fluorescens cumene dioxygenase PCR

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