Pseudomonas aeruginosa has variety of virulence factors that contribute to its pathogenicity. Therefore, rapid detection with high accuracy and specificity is very important in the control of this pathogenic bacterium. To evaluate the accuracy and specificity of Polymerase Chain Reaction (PCR) assay, ETA and gyrB genes were targeted to detect pathogenic strains of P. aeruginosa. Seventy swab samples were taken from patients with infected wounds and burns in two hospitals in Erbil and Koya cities in Iraq. The isolates were traditionally identified using phenotypic methods, and DNA was extracted from the positive samples, to apply PCR using the species specific primers targeting ETA, the gene encoding for exotoxin A, and gyrB gene. The results of this study indicate that 100% of P. aeruginosa isolates harbored the gyrB gene, whereas 74% of these isolates harbored ETA gene. However, the specificity of PCR for detection of P. aeruginosa based on the both genes was 100%, since no amplified product obtained using DNA extracted from other bacterial species. Hence by considering the importance of rapid detection of this bacterium due to the presence of problems in biochemical methods, PCR targeting multiple virulence genes is suggested in identification of pathogenic strains of P. aeruginosa isolated from some infections which should speed diagnosis of an antimicrobial therapy.
As major nosocomial pathogens,
In this study, 20
Staphylococcus aureus and Pseudomonas aeruginosa are the major globally distributed pathogens, which causes chronic and recalcitrant infections due to their capacity to produce biofilms in large part. Biofilm production represents a survival strategy in these species, allowing them to endure environmental stress by altering their gene expression to match their own survival needs. In this study, we co-cultured different clinical isolates of S. aureus and P. aeruginosa as mono- and mixed-species biofilms in a full-strength Brain Heart Infusion Broth (BHI) and in a 1000-fold diluted Brain Heart Infusion Broth (BHI/1000) using Microtiter plate assay and determination of colony-forming units. Furthermore, the effect of starvation stress on the e
... Show MoreThis study ,the samples were collected from "118 patients " suffering from burn wound contaminated with Pseudomonas aeruginosa and 100 health individuals (male and female ) as a control group ,the samples were wound swap and blood sample . Chromatography technique was employed to extract and purify cell wall containing lipopolysaccharide by using P. aeruginosa isolate ATCC 15692,the purification done by addition of ammonuium sulfate, sodium dodecyl sulfat (SDS) anddialysis, gel filtration chromatography by using sepharose-4B. Immunogenicity of LPS component was determined by mice injection under the skin ,then Ab concentration agai
... Show MorePseudomonas aeruginosa was isolated from various clinical samples included urine, sputum, stool, ear, wound & burn swabs. Detection of the ability of local isolates to produce staphylolysin enzyme was studied, on Tryptic soya agar + 0.2% (wt./vol.) of heat killed Staphylococcus. aureus at temperature 100oC. medium and the diameters of lysis zone ranged from 5-22mm, then the isolate P16 was chosen to extract staphylolysin A (LasA) and its specific activity reaches 8.59 unit /mg protein, whi1e the isolate P5 was chosen to extract staphylolysin D (LasD) where it's specific activity reaches 0.66 unit /mg protein since the two isolates were the most production of enzyme. Staphylolysin enzyme was extracted by cooling centrifugation and par
... Show MoreThe present study aims to evaluate the synergistic activity of nicotinic acid (NIC) with the Imipenem (IMI) as an anti-biofilm for clinical isolated Pseudomonas aeruginosa. The values of minimum inhibitor concentration (MICs) for IMI and NIC (Separately) against P. aeruginosa were (16) ug/mL and (8) ug/ml respectively. Whereas, the concentration of NIC with IMI (as combined) for biofilm inhibition was 1 ug/ml for NIC and 4 ug/ml for IMI. The combining of NIC with IMI showed synergistic efficacy against formation of bacterial biofilm (at MIC levels). These results provide a conclusion that NIC combined with IMI is can be considered as a successful prospective treatment against the biofilm pr
... Show MoreThe study was aimed at inhibiting the protease produced by Pseudomonas aeruginosa using an 80% alcoholic extract of Conocarpus lancifolius leaves. A total of 146 isolates of P. aeruginosa that were isolated and identified by microscopic and biochemical tests were 51 isolates submitted to primary and secondary screening techniques in order to choose the qualified P. aeruginosa isolate for protease synthesis. Among these isolates, forty-seven isolates showed hydrolysis zones on skim milk media (primary screening); six isolates were chosen for secondary screening. The result revealed that P. aeruginosa P51 had the highest ability to produce the enzyme, with a specific activity of 15.9 U/
... Show MoreNine Iraqi varieties of barley (Hordeum vulgare L.) has been differentiated and diagnosed using simple sequence repeat markers to detect their genetic polymorphism. Six SSR primers were used for genetic screening of barley samples (IPA 265, IPA 99, Tuwaitha, Hitra, Rayhan, Shuaa, Bawadi, Samir and Al_khair). These primers generated total PCR product (11) bands divided to 8 polymorphic bands 3 monomorphic bands. the percentage of polymorphism 80% ranged between (50-100%). a mean value of polymorphic band per primer was 1.6 . these primers produced amplification fragment at Molecular weight between 75-900 bp. One unique band was generated at size 200bp, this band can be used as a DNA profiling of all studied genotypes. These results appear
... Show MoreGenetic variation was studied in 22 local and imported samples collected from local Iraqi market by using random amplified polymorphic DNA (RAPD-PCR). Five randomly primers set were used in this study. These primers produced 292 bands. Molecular weights of these bands ranged between 1.8 Kb (1800 bp) to 150 bp. The percentage of polymorphic bands is 100%, with one distinguished band which is produced by using C52 primer. The other primers did not produce any distinguished band. The results of Dendrogram of the studied samples depended on RAPD-PCR results by using Jaccard coefficient for genetic similarity was distributed the samples into 8 groups. This Dendrogram revealed a higher similarity between Iraqi/Yousifia green bell pepper and Jo
... Show MorePseudomonas aeruginosa is a common and major opportunistic human pathogen, its causes many and dangersinfectious diseases due to death in some timesex: cystic fibrosis , wounds inflammation , burns inflammation , urinary tract infection , other many infections otitis external , Endocarditis , nosocomial infection and also causes other blood infections (Bacteremia). thereforebecomes founding fast and exact identification of P. aeruginosafrom samples culture very important.However, identification of this species may be problematic due to the marked phenotypic variabilitydemonstrated by samples isolates and the presence of other closely related species. To facilitate species identification, we used 16S ribosomal DNA(rRNA) sequence data
... Show MoreOwing to high antibacterial resistance of Pseudomonas aeruginosa, it could be considered as the main reason behind the nosocomial infections. P. aeruginosa has a well-known biofilm forming ability. The expression of polysaccharide encoding locus (pelA gene) by P. aeruginosa is essential for this ability. The purpose of the current research was to determine the biofilm formation in P. aeruginosa isolated from clinical samples and to evaluate the role of the selected PelA gene in biofilm formation using PCR method in Iraqi patients. Results revealed that 24 (96%) isolates were found to have the ability to form biofilm that was remarkably related to gentamicin resistance. Moreover, the pelA gene was found in all biofilm-producers. In conclu
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