Ümit KILIÇ, Tayfur DEMİRAY, Mustafa ALTINDİŞ

KARBAPENEMAZ ÜRETEN ENTEROBACTERIACEAE İZOLATLARININ SAPTANMASINDA FENOTİPİK VE GENOTİPİK METOTLAR

Başta karbapenem direnci olmak üzere çoklu ilaç direnci hem hastane hem de toplum kaynaklı salgınlara neden olmakta, dirençli bakteri türlerinin çeşitliliğini artırmakta ve tüm dünyada alarm düzeyinde hızla yayılmaktadır. İnfekte hastalar ve kolonize taşıyıcıların erken ve etkin şekilde saptanması, çoklu ilaç direncininin yayılmasının önlenmesi ve infeksiyon kontrolünde zorunlu adımdır. Güncel EUCAST rehberi karbapenemaz üreten Enterobacteriaceae'nın belirlenmesinde, direncin maksimum duyarlılıkta saptanmasını garantilemek için pozitif örneklerde düşük klinik sınır değerlerinin kullanılmasını önermektedir. Günlük iş akışı içinde direnç paterni ve türünü saptamak için öncelikle bir tarama yöntemi gereklidir, ardından bunu fenotipik ve/veya genotipik olarak bir doğrulama testi takip edebilir.Burada kullanılabilecek fenotipik testler; Modifiye Hodge Testi (MHT), inhibitör bazlı yöntemler (çift disk sinerji testi, kombine disk testleri, E test-MBL vb.), kromojenik besiyerleri, biyokimyasal yöntemler (Carba NP, Blue carba), MALDI-TOF MS ve immünokromotojenik yöntemler iken başlıca genotipik metotlar ise PZR çeşitleri, oligonükleotid hibridizasyonu, PFGE ve MLST şeklindedir. Karmaşık direnç paternli patojenleri saptamak ve yayılımı kontrol etmek için hem optimize fenotipik testler hem de genotipik analizler klinik laboratuvarlarının rutin tanısal testleri olarak önerilmektedir

Phenotypic and Genotypic Methods for Determination of Carbapenemase Producing Enterobacteriaceae Isolates

Multidrug resistance, particularly, carbapenem resistance is spreading worldwide at an alarming rate, comprehending a variety of bacterial species and causing both nosocomial and community acquired outbreaks. Early and efficient detection of infected patients or colonized carriers is the mandatory step in infection control and prevention of multidrug resistance spread. The latest EUCAST guidelines for detection of carbapenemase-producing Enterobacteriaceae have proposed using low clinical breakpoints to ensure the maximum detection sensitivity of positive samples. Current workflows involve an initial screening step for species and resistance pattern detection, followed by phenotypic and/or genotypic confirmation. Phenotypic tests for determination of the carbapenemase-producing Enterobacteriaceae were Modified Hodge Test (MHT), inhibitor-based methods (double-disk synergy test, combined disk test, E Test-MBL), chromogenic culture media, biochemical methods (Carba NP, Blue carba etc.), MALDI-TOF MS, immunochromotogenic methods; the genotypic methods were PCR, oligonucleotide hybridization, PFGE ve MLST. To detect and to control the spread of pathogens with complicated resistance patterns, both optimized phenotypic analysis and genotypic assays are recommended in the routine diagnostic of clinical laboratories

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