CANDIDA ALBICANS’ A AİT ALS, HWP VE STAPHYLOCOCCUS EPIDERMIDIS’ E AİT ICA GENLERİNİN FARKLI BİYOMATERYALLERDEKİ EKSPRESYONLARININ REAL TİME PCR İLE KANTİTATİF ANALİZİ

Çalı!mamızda lateks-slikon, polivinil klorid, polivinilprolidon kaplı poliüretan ve poliüretan kateter yüzeyinde olu!turulan in vitro biyofilm modelinde adezyon genlerinin ekspresyonları kar!ıla!tırılmı!tır. Kar!ıla!tırılan adezyon genleri sırasıyla Candida albicans \'a ait ALS1 ve HWP1, Staphylococcus epidermidis \'e ait ICA \'dır. Olu!an biyofilm kütlesi kateter yüzeyinden kazınmı! ve mRNA \'ları izole edilmi!tir. Bu mRNA \'lar cDNA \'ya çevrilmi! ve real time PCR \'da kantitatif analizleri yapılmı!tır. Çalı!mamızın sonunda slikonize materyalden yapılmı! kateter üzerindeki ALS1 ve HWP1 genlerinin kopya sayısı sırasıyla 2,50 x 105 kopya/ml ve 1,30 x 104 kopya/ml, ICA geninin ise 1,80 x 105 kopya/ml oldu\"u bulunmu!tur. Adezyon genleri polivinil klorid kateterde slikonize olanlardan daha çok eksprese edilmi! olarak bulunmu!tur. Polivinil klorid kateter üzerindeki ALS1 ve HWP1 genlerinin kopya sayısı sırasıyla 7,56 x 106 kopya/ml ve 6,68 x 105 kopya/ml olarak, ICA geninin ise 4,20 x 105 kopya/ ml olarak bulunmu!tur. Minimum gen ekspresyonu polivinilprolidon kaplı poliüretan ve poliüretan kateterlerdede saptanmı!tır. Biyofilm ili!kili genler PVP ve PUC yüzeyinde veya planktonik hücrelerde farklı bulunmamı!tır. Bu çalı!mada, adezyonla ili!kili genlerin ekspresyonunun materyalin yapısından etkilendi\"i bulunmu!tur. Bu klinik kullanımda, bir materyalin di\"erine tercih edilmesiyle sonuçlanabilir.The Quantitation and Comparison of gene expression levels in different biomaterials for ALS1 and HWP1 in Candida albicans, for ICA in Staphylococcus epidermidis in Real Time PCR. This study describes a comparison of latex-silicone coated, polyvinyl chloride, polyvinylpyrolidone coated polyurethane and polyurethane catheters for the delivery of adhesion genes in an in vitro biofilm model. ALS1, HWP1 genes of Candida albicans and ICA gene for Staphylococcus epidermidis were targeted for the comparison of four different biomaterials. Biofilm mass was separeted from the surface of the catheter and mRNA of the strains were isolated. Isolated mRNA was transcripted into cDNA and quantified in a real time PCR assay. As a result of our study, ALS1 and HWP1 gene copy numbers were found to be 2.50x105 and 1.30x104 copy/ml, respectively and ICA gene copy number was found to be 1.80x105 copy/ml on the biofilm of the catheter made from siliconized material. Adhesion genes were found as expressed in higher levels on the polyvinyl chloride catheter than those in siliconized ones. ALS1 and HWP1 gene copy numbers were found to be 7.56x106 and 6.88x105 copy/ml, respectively and ICA gene copy number was found to be 4.20x105 copy/ml on the polyvinyl chloride catheter. Minimum gene expression levels were detected in polyvinylpyrolidone coated polyurethane and polyurethane catheters. Biofilm related genes were not expressed differently between planktonic cells or on the surfaces of those catheters. In this study, the expression of the adhesion related genes were found to be affected from the structure of the materials. It may result in the preferation of one material from other in the clinical use

Investigating Hemolytic Activity of Candida Isolates with Two Different Methods

The hemolytic activity of Candida isolates with agar and microplate methods were investigated and compared efficiency of these methods to assess relationship between hyphal formation and hemolysis