Esra Deniz CANDAN, Nilüfer AKSÖZ, Işıl Seyis BİLKAY

RAPD-PCR yöntemi ile metisilin duyarlı ve dirençli Staphylococcus aureus klinik izolatları arasındaki genetik ilişkinin saptanması

M etisilin dirençli Staphylococcus aureus MRSA görülme sıklığındaki artış bu suşların karakterizasyonunun hızlı ve güvenilir bir şekilde yapılmasını ve hastanelerde klonal yayılımın tanımlanmasını gerektirmektedir. Bu çalışmanın amacı, klinik örneklerden elde edilen Staphylococcus aureus suşlarına ait rastgele çoğaltılmış polimorfik DNA RAPD bant modellerindeki varyasyonu ve bunlar arasındaki sistematik ilişkiyi ortaya çıkarmaktır. Suşların tanımlanması ve antibiyotik duyarlılıkları otomatize sistem ve geleneksel yöntemler ile belirlendi. Toplam 47 izolatın 30’u %64’ü metisiline dirençli bulundu. Metisilin duyarlı ve dirençli izolatlar karşılaştırıldığında metisilin dirençli izolatlarda en düşük etki gösteren antibiyotikler rifampin, tüm aminoglikozidler ve penisilinler, tetrasiklin ve amoksilin-klavulonik asittir. 47 S.aureus klinik izolat, RAPDPCR tekniği kullanılarak 25 farklı primer ile analiz edildi. Tüm veriler ile ağırlıksız çift grup yöntemi UPGMA kullanılarak dendogram çizildi. Bu çalışmada, beş primerin kombinasyonu kullanılan RAPD-PCR yöntemi klinik örneklerden izole edilen MRSA ve MSSA izolatları arasındaki genetik ilişkinin belirlenmesinde başarılı bir şekilde uygulanabildiği açıkça görülmektedir

Anahtar Kelimeler:

Antibiyotipleme, Staphylococcus aureus, Metisilin direnci, RAPD-PCR

Determination of genetic relationship between methicillin-sensitive and methicillin- resistant Staphylococcus aureus clinical isolates by RAPD-PCR Method

The increase in the frequency of methicillin-resistant Staphylococcus aureus MRSA demands a quick and trustworthy characterization of isolates and identification of clonal spread within hospitals. The objective of this study is to establish variations of randomly amplified polymorphic DNA RAPD patterns in S. aureus isolates which are collected from clinical isolates and a systematic relationship between them. Identification of isolates and antimicrobial susceptibility tests were performed by automated system and conventional phe- notypic methods. A total of 30 out of 47 isolates 64% expressed methicillin-resistance. When methicillin-sus- ceptible and resistance isolates were compared, the least active agents were rifampin, all aminoglycosides and penicillines, tetracycline and amoxicillin-clavulonic acid for methicillin-resistance isolates. RAPD-PCR method with 25 different primers was analyzed on 47 clinical isolates of S.aureus. All the data were employed to cons- truct dendograms using the unweighted pair group method using arithmetic UPGMA . In this study, it is clear that, RAPD-PCR assayed with combination of five primers can be successfully applied to assess the genetic relationship between MRSA and MSSA isolates from clinical samples.

Keywords:

Antibiotyping, Staphylococcus aureus, methicillin resistance, RAPD-PCR,

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