[Evaluation of exotoxin A gene and frequency of polymerase chain reaction sensitivity in detection of pseudomonas aeruginosa isolated from burn patients]

Background: Pseudomonas aeruginosa is a gram-negative pathogens opportunism which causes severe infections in human beings

The most common infection include: endocarditis, meningitis, septicemia and chronic lung infections in cystic fibrosis patients

This bacterium has many pathogenic factors including; exotoxin A, lipopoly-sacharide, phospholipase C, pili, elastase and alkaline protease. The purpose of this study was to evaluate the frequency of exotoxin A gene [ETA] as a strong virulence factor and sensitivity determination of polymerase chain reaction [PCR] in pseudomonas aeruginosa isolated from second and third-degree burn patients

Methods: This study has performed in Besat University Hospital in Hamadan from January to December 2012. We used 170 isolated samples

The samples were isolated from blood and skin biopsy in second and third-degree burn patients

We had 79 strains positive culture of pseudomonas aeruginosa. Forward and reverse primers used for PCR were designed by DNASIS and Oligo software. Then genomic of known strains were extracted by DNA purification kit and indemnified by PCR. The quality and quantity of the extracted DNA was determined using spectrophotometry

For determination of PCR sensitivity was used culture test as gold standard. DNA of pseudomonas aeruginosa [ATCC 27853] was used as a positive control. Finally data was analyzed using SPSS software

Results: Out of 170 isolated samples, 79 strains of pseudomonas aeruginosa isolated from burn patients had positive culture

PCR of isolated positive culture demonstrated that 5 strains [6.33%] were with out this virulence factor and 74 strains [93.67%] had ETA gene. So the sensitivity of test based on sensitivity formula was 94.04%. Conclusion: Our results showed that sensitivity of PCR mediated ETA gene in detection of pseudomonas aeruginosa strains is considerable and this factor can be used as a good factor identifying of pseudomonas aeruginosa. It seems more studies with larger sample size is necessary in this area

[Association between cholestryl ester transfer protein D442G polymorphism on serum lipid levels and CETP activity in hypercholesterolemic patients]

Hypercholesterolemia is considered a major risk factor for pancreatitis, atherosclerosis and coronary heart disease. Cholesteryl ester transfer protein gene polymorphisms are known to be associated with changes in lipid levels. We investigated the association between a polymorphism in the CETP gene [D442G] with plasma lipid levels and CETP activity in patients with hypercholesterolemia. This case/control study that be done in Hamadan university of medical sciences [from October 2008 to September 2009], included 102 patients with hypercholesterolemia and 200 healthy individuals. Polymerase chain reaction and restriction fragment length polymorphisms were used to determine genotypic distribution and allelic frequencies of polymorphisms. The plasma CETP activity was measured by a kit in a fluorescence spectrometer. Lipid concentrations were measured by routine biochemical and enzymatic assays. Plasma cholesteryl ester transfer protein activity was significantly higher in the cases than the controls [P<0.05]. The genotypic and allelic frequencies for this polymorphism were not statistically different between the patients with hypercholesterolemia and the controls [in controls: DD 96%, DG 4%, GG 0% and in cases: DD 86%, DG 10%, GG 4%], [P>0.05]. Plasma HDL-C, LDL-C and TC were higher in both groups with GG and DG genotypes than with DD genotype, whereas serum CETP activity was lower in GG genotype compared with other genotypes [GD or DD], [P<0.05]. The results showed that D442G polymorphism of CETP gene was associated with changes in lipid profile and plasma CETP activity in the selected population and it might have a role in contributing to a genetic risk for developing coronary artery disease