Staphylococcus aureus izolatlarının tiplendirilmesinde çoklu lokus değişken sayı tekrar analizi (MLVA) ve darbeli alan jel elektroforez (PFGE) yöntemlerinin karşılaştırılması

Sümeyra Savaş

Öz


Bu çalışmada, Staphylococcus aureus (S. aureus) bakterisi için, çoklu lokus değişken sayı tekrar analizi (MLVA) yönteminin, altın standart olarak bilinen moleküler tiplendirme yöntemi darbeli alan jel elektroforezi (PFGE) ile ayırt edebilme performansı araştırıldı. S. aureus için PFGE yönteminden elde edilen sonuçların MLVA yöntemi ile ne oranda sağlanabileceği belirlenmeye çalışıldı. Bu sayede rutin kullanımlarda zorlu bir yöntem olan PFGE yönteminin, daha kolay ve düşük maliyetli bir yöntem olan MLVA ile rutin de yer değiştirmesinin olanağı araştırıldı.
MLVA yöntemi hızlı bir DNA izolasyon yöntemi sonrası, sdrCDE, clfA, clfB, sspA ve spa lokusları için polimeraz zincir reaksiyonu (PCR) yöntemi kullanılarak gerçekleştirildi. PFGE yöntemi ise SmaI enzimi kullanılarak elde edilen genomik DNA fragmanları ile gerçekleştirildi. Bantlama modeli, hem görsel olarak hem de Bionumerics yazılımı ile analiz edildi. PFGE ile izolatlar 2 küme ve 16 alt tipe ayrılırken, MLVA ile aynı izolatlar, küme oluşturmaksızın 31 alt tipe ayrıldı. PCR temelli bu yöntem ile, PFGE yöntemi ile ayırt edilemeyen alt tiplerin ayırt edilebildiği görüldü. Bu sebep ile MLVA yönteminin aynı PFGE paternleri ile ilgisiz suşları daha iyi tanımlamak veya ilişkili izolatların yakın genetik kompozisyonunu doğrulamak için yüksek epidemiyolojik çözünürlüğe sahip, alternatif, verimli moleküler tipleme yöntemi olarak kullanılabileceği sonucuna varıldı.


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