Detection of Carbapenem Resistance Using the Genotypic and Phenotypic Methods in Klebsiella pneumoniae

Abstract

Aim: This study aimed to detect the carbapenem resistance of the Klebsiella pneumoniae strains, isolated from clinical specimens with genotypic and phenotypic methods. Material and Methods: A total of 87 Klebsiella pneumoniae strains whose carbapenem resistance was determined by disc diffusion method were included in the study. Carbapenemase was investigated using the combined disk method and polymerase chain reaction (PCR). Results: The evaluation of the PCR results demonstrated that OXA was detected in 60 (68.9%) samples, NDM was detected in 20 (22.9%), OXA + NDM in 5 (5.7%), and KPC was detected in 1 (1.1%) out of 87 clinical samples. Carbapenemase was not detected in one specimen with the PCR method. The results were found compatible with the combined disc test results for all isolates which were detected as only OXA, NDM, and KPC type carbapenemase positive. In 5 (5.7%) strains in which the co-existence of NDM and OXA type carbapenemases was detected by PCR, the combined disc method detected only OXA type carbapenemase. Conclusion: The combined disk method is inadequate in the presence of strains that have multiple carbapenemases, and also have OXA which is the most frequently detected carbapenemase in our hospital. EUCAST recommends verification by other methods in the presence of OXA-48. Genotypic methods can be used for confirmation testing. The detections of strains with NDM, multiple carbapenemases, and the first detection of KPC were striking in the study. Monitoring the spread of these strains in the hospital will be necessary for infection control.

Keywords

• Klebsiella pneumoniae
• carbapenem
• carbapenemase

Klebsiella pneumoniae Suşlarında Karbapenem Direncinin Genotipik ve Fenotipik Yöntemler ile Saptanması

Abstract

Amaç: Bu çalışmada, klinik örneklerinden izole edilen Klebsiella pneumoniae suşlarında karbapenem direncinin genotipik ve fenotipik yöntemler ile saptanması amaçlanmıştır. Gereç ve Yöntemler: Disk difüzyon yöntemi ile karbapeneme dirençli bulunan toplam 87 Klebsiella pneumoniae suşu çalışmaya dahil edilmiştir. Karbapenemaz varlığı kombine disk yöntemi ve polimeraz zincir reaksiyonu (polymerase chain reaction, PCR) ile araştırılmıştır. Bulgular: PCR sonuçları değerlendirildiğinde, 87 klinik örnekten 60 (%68,9) örnekte OXA tipi, 20 (%22,9) örnekte NDM tipi, 5 (%5,7) örnekte OXA + NDM tipi ve 1 (%1,1) örnekte ise KPC tipi karbapenemaz saptandığı görülmektedir. Bir örnekte ise PCR yöntemi ile karbapenemaz bulunamamıştır. Tek başına OXA, NDM ve KPC tipi karbapenemaz pozitifliği saptanan izolatların tamamı için sonuçların kombine disk testi ile uyumlu olduğu bulunmuştur. PCR yöntemi ile NDM ve OXA tipi karbapenemazın birlikte olduğu 5 (%5,7) örnekte ise kombine disk yönteminde sonuç OXA tipi karbapenemaz olarak bulunmuştur. Sonuç: Kombine disk yöntemi, aynı anda birden fazla karbapenemaz bulunduran suşların bulunması ve hastanemizde en sık saptanan karbapenemaz tipinin OXA olması nedeni ile yetersiz kalmaktadır. EUCAST, OXA-48 varlığında diğer yöntemlerle doğrulanmasını önermektedir. Doğrulama testi olarak genotipik yöntemler kullanılabilir. NDM tipi karbapenemazın artmakta olduğu, birden fazla karbapenemazı taşıyan suşların görülmeye başlaması ve ilk defa KPC tipi karbapenemazın bulunması dikkat çekicidir. Bu suşların hastanede yayılımının takip edilmesi enfeksiyon kontrolü açısından önemli olacaktır.

Keywords

• Klebsiella pneumoniae
• karbapenem
• karbapenemaz

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