Tek ve iki türlü biyofimlerde E.faecalis jelatinaz gen ekspresyon seviyelerinin belirlenmesi

Year 2015, Issue: 2, 162 - 173, 01.06.2015

Didem Kart

Abstract

Biyofilmler insan hayatını tehdit eden önemli bir problemdir. Son yıllarda, persistan enfeksiyonlara yol açan biyofilmlerin çoğunun polimikrobiyal olduğu gösterilmiştir. Enterococcus faecalis idrar yolu, yara enfeksiyonları vb. gibi hastane enfeksiyonlarından sıklıkla izole edilen fırsatçı bir patojendir. E.faecalis’in bazı virulans faktörlerinin ekspresyon seviyelerinde, biyofilm oluşturabilen ve oluşturamayan izolatları arasında farklılıklar gözlenmiştir. Jelatinaz, jelatini, kollajeni ve kazeini hidrolize eden metallo-proteaz enzimi olup, E.faecalis’in virulans faktörlerinden birisidir. Mutant suşlar kullanılarak yapılan bazı çalışmalarda bu enzimin biyofilm oluşumundaki rolü bulunmuştur. İlk amacımız, 11 E.faecalis idrar izolatının biyofilm oluşturabilme prevalansını belirlemektir. İkinci amacımız, tekrar edilebilirliği yüksek iki farklı mikrobiyal türden oluşan biyofilm modelini in vitro ortamda geliştirmek ve C.albicans’ın ortamdaki varlığının E.faecalis’in gelE gen ekspresyonu üzerindeki etkisinin kantitatif RT-PCR ile belirleyebilmektir. Çalışmamızda, test edilen her iki yöntemde de tüm izolatların biyofilm oluşturabildiği saptanmıştır. Ayrıca, E.faecalis ATCC 29212 suşu tarafından oluşturulan monomikrobiyal biyofilm modeline oranla, C.albicans ATCC 90028 ve E.faecalis ATCC 29212 tarafından oluşturulan ikili biyofilm modelinde gelE ekspresyon artışı olduğu sonuçlarımızda gösterilmiştir. İki türden oluşan biyofilm modelinde gözlenen bu gelE ekspresyon seviyesindeki artışı desteklemek amacıyla klinik izolatlar kullanılarak yapılacak çalışmalara ihtiyaç duyulmaktadır.

Keywords

Biyofilm, E.faecalis, Jelatinaz geni, Polimikrobiyal, Ekspresyon

Determination of Gelatinase gelE Expression Levels of Enterococcus faecalis in Mono and Dual Species Biofi lms

Abstract

Biofilms are substantial problem threatening human health. Recently, it was shown that most of biofilms leading to persistent infections were polymicrobial. E. faecalis is an opportunistic pathogen frequently isolated from hospital infections such as urinary tract, wound infections etc. Differences on expression levels of some virulence factors of E.faecalis were determined between biofilm producing and nonproducing isolates. Gelatinase, a metallo protease enzyme hydrolyzing gelatin, collagen and caseine, is one of the virulence factors of E.faecalis. The role of gelatinase with biofilm formation was found in some studies using mutant strains. Our first aim is to determine the prevalence of biofilm forming ability of 11 E.faecalis urine isolates. Our second aim is to set up an in vitro dual biofilm model in a repeatable style and to find out the impact of the presence of C.albicans on the gelE gene expression of E.faecalis by using RT-qPCR. In the present study, all isolates were found as biofilm producer by both method. Moreover, our results showed increased gelE expression in dual species biofilm formed by C. albicans ATCC 90028 and E. faecalis ATCC 29212 when we compared with E. faecalis monomicrobial biofilm form. Further studies with clinical isolates will be needed to support this increased gelE expression level in the dual biofilm.

Keywords

Biofilm, E.faecalis, Gelatinase gene, Polymicrobial, Expression

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