Reyhan YİŞ, E. Deniz BAYRAM, Özlem Yüksel ERGİN

BLAOXA-48 POZİTİF K. PNEUMONİAE İZOLATLARINDA FENOTİPİK KARBAPENEMAZ ÜRETİMİNİN MODİFİYE HODGE VE KARBAPENEMAZ İNAKTİVASYON TESTLERİ İLE DEĞERLENDİRİLMESİ

Amaç: Son yıllarda özellikle uzun süre hastanede yatan, immun supresif, invaziv araç ve geniş spektrumlu antibi¬yotik kullanılan hastalarda kolonizasyon oranları gittikçe artmakta olan K.pneumoniae, dirençli enfeksiyonlara yol açmaktadır. Çalışmamızda laboratuvarımıza gönderilmiş olan klinik örneklerden izole ettiğimiz çoklu dirençli K. pneumoniae izolatlarında karbapenem direncinin fenotipik ve genotipik yöntemlerle saptanması amaçlanmıştır. Materyal ve Metot: Temmuz 2014’te çeşitli servislerden labora¬tuvarımıza gelen örneklerden izole edilen, 51 K. pneumoniae değerlendirilmiştir. İzolatların tür düzeyinde tanımlanması ve antibiyotik duyarlılık testleri otomati¬ze sistem (BD Phoenix System, ABD) ile çalışılmıştır. Karbapenem direnci gradient test stripleri (BioMerieux, Fransa ve Liofilchem, İtalya) ile doğrulanmıştır. Karbapenemaz direnç genlerinin saptanması ‘in-house’ polimeraz zincir tepkimesi (PZT) ile gerçekleştirilmiştir. Tüm izolatlara fenotipik testlerden Modifiye Hodge testi (MHT) ve Karbapenemaz İnaktivasyon Testi (CIM Testi) uygulanmıştır. MHT ve CIM testi uyumsuz sonuç veren izolat için her iki test tüm karbapenemler ile tekrar çalışılmıştır. Bulgular: İmipenem (IPM), meropenem (MEM) ve ertapenem (ETP) direnç oranları sırasıyla %78,43, %60,78 ve %96,08 olarak belirlenmiştir. MHT ile 51 adet izolattan 42 ‘sinde, CIM testi ile 43 adet izolatta pozitiflik saptanmıştır. PZT sonuçlarına göre izolatların 42‘sinde blaOXA-48 geni bulunduğu belirlenmiş olup, diğer karbapenemaz genleri saptanmamıştır. blaOXA-48 geni ve diğer karbapenemaz genleri negatif olarak saptanan dokuz izolatın tamamında MHT, sekiz izolatta ise CIM Testi ile negatiflik saptanmıştır. Sonuç: MHT ve CIM, karbapenemaz varlığının saptanmasında basit ve hızla uygulanabilecek yöntemlerdir. CIM, değerlendirilmesi daha kolay, bir tarama testi olarak faydalı olabilecek bir yöntem olsa da, yalancı negatif ve pozitif sonuçlar açısından dikkatli olmayı gerektirmektedir. Testin diğer karbapenem disklerinin kullanımı yoluyla modifiye edilmesinin yalancı negatiflikleri azaltabileceğini düşünüyoruz.

Evaluation of Phenotypic Carbapenemase Production in blaOXA-48 positive K. pneumoniae Isolates by Modified Hodge and Carbapenemase Inactivation Tests

Aim: In recent years, increasing colonization rates of K. pneumoniae in patients with immunosuppression, long hospital stay, invasive devices and treated with broad spectrum antibiotics lead to resistant infections. In this investigate, we aimed to investigate carbapenem resistance in multiresistant K. pneumoniae isolates derived from clinical specimens by phenotypic and genotypic methods. Materials and Methods: In July 2014, 51 K. pneumoniae isolates which were derived from clinical specimens were evaluated. Identification and antibiotic susceptibility tests of isolates were studied with automatized system. Verification of carbapenem resistance were studied with with gradient strip tests. Identification of carbapenemase resistant genes were studied with with “in house” PCR. Phenotypic tests including Modified Hodge test (MHT) and Carbapenemase Inactivation Method (CIM) were applied to all isolates. We re-studied all carbapenems for the isolates with incompatible results. Results: Resistance rates of all isolates to imipenem, meropenem and ertapenem were 78.43%, 60.78% and 96.08%, respectively. 42 of 51 isolates were positive with MHT and 43 of 51 isolates were positive with CIM. blaOXA-48 gene was identified in 42 isolates with PCR. nine isolates which were negative for blaOXA-48 and other carbapenemases gene were negative with MHT and eight isolates which were negative for blaOXA-48 gene were negative with CIM. Conclusion: MHT and CIM are fast and easy methods for the identification of carbapenemase presence. CIM is an easy screening test, but one should be cautious for false negative and positive results. We suggest that modification of the test with carbapenem discs should decrease false negative results.

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