* Bu çalışma, 3. Ulusal KLİMİK Kongresi (18-22 Kasım 2015, Antalya)'nde sunulmuştur. ÖZEnfeksiyonların uygun olarak tedavi edilebilmesi ve enfeksiyon kontrol önlemlerinin uygulanabilmesi için karbapenemaz üreten Enterobacteriaceae izolatlarının hızlı, pratik ve doğru bir şekilde tanımlanması çok önemlidir. Bu amaçla çok sayıda fenotipik test yöntemi geliştirilmiş olmasına rağmen, hiçbirisi %100 duyarlılık ve özgüllüğe sahip değildir. Testlerin duyarlılık ve özgüllüğü, çalışılan izolatların taşıdığı betalaktamaz enzim tiplerine göre değiştiğinden, ülkeler ve bölgeler arasında farklılıklar görülebilmektedir. Bu çalışmada, Enterobacteriaceae ailesi üyelerinde karbapenemazların saptanmasında yaygın olarak kullanılan modifi ye Hodge testi (MHT) ve "Carbapenemase Nordmann Poirel (Carba NP)" testlerinin performanslarının karşılaştırılması amaçlanmıştır. Çalışmaya çeşitli klinik örneklerden izole edilen ve en az bir karbapeneme (ertapenem, imipenem veya meropenem) azalmış duyarlılık gösteren ve karbapenemaz geni taşıdığı önceden polimeraz zincir reaksiyonu (PCR) ile saptanan 65 Enterobacteriaceae izolatı (43 bla OXA-48 , 10 bla VIM , 9 bla IMP , 1 bla NDM-1 , 1 bla KPC-2 ve 1 bla OXA-48 +bla VIM taşıyan suş) dahil edilmiş-tir. Karbapenemaz geni saptanmayan fakat karbapenemlere azalmış duyarlılık gösteren kontrol grubu olarak 78 izolat kullanılmıştır. Tüm izolatlar konvansiyonel yöntemlere ilaveten BD Phoenix otomatize sistemi (Becton Dickinson, ABD) ile tanımlanmıştır. İzolatların antimikrobiyal duyarlılıkları, BD Phoenix otomatize sistemi ile saptanmış ve disk difüzyon yöntemi ile doğrulanmıştır. Sonuçlar CLSI kriterlerine Geliş Tarihi
The rapid and accurate identification of carbapenemases is of crucial importance in terms of infection control. Methods employed in the determination of carbapenemases should be constantly updated in the light of technical advances and newly emerging carbapenemase variants. The aim of this study was to evaluate the performance of the newly developed carbapenem inactivation method (CIM) for the identification of carbapenemases defined in the members of the family Enterobacteriaceae. Enterobacteriaceae isolates with resistance to at least one of the carbapenems (ertapenem, imipenem or meropenem) were included in the study. The study isolates were obtained from various clinical specimens between 2008-2014 and consisted of 56 Enterobacteriaceae strains (12 Escherichia coli, 32 Klebsiella spp., and 12 Enterobacter spp.) in which the presence of the 38 blaOXA-48, 8 blaVIM, 7 blaIMP, 1 blaNDM-1, 1 blaKPC-2 and 1 blaOXA-48+blaVIM genes had been previously determined using the polymerase chain reaction (PCR) and 78 in which no carbapenemase gene were detected. For the performance of the CIM, the test bacteria were suspended in sterile water and then a 10 μg meropenem disc was immersed in the suspension and incubated for 2 hours. This meropenem disc was then removed and subsequently placed on a Mueller-Hinton agar plate inoculated with E. coli ATCC 29522 and incubated at 35°C. The results were assessed after 6 hours and after overnight incubation. Development of an inhibition zone around the meropenem disk was interpreted as the absence of carbapenemase and the lack of an inhibition zone as the presence of carbapenemase. The results of the CIM were obtained after 8 hours. With the CIM, all isolates with previously determined carbapenemase genes were found to be positive and the isolates with no genes revealed to be negative. The sensitivity and specificity of CIM were estimated as 100%. The high sensitivity and specificity, ease of application and interpretation, rapid production of results, and no necessity for additional equipment or chemical substances, makes CIM a promising high throughput method to be used in routine microbiology laboratories. Since this method indicates only the presence of a carbapenemase in a clinical isolate, the type of the carbapenemase present should be further identified by the use of molecular techniques.
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