Yoğun bakım ünitelerinde kan dolaşımı enfeksiyonu etkeni karbapenem dirençli Acinetobacter baumannii izolatlarının moleküler yöntemlerle karakterizasyonu

Year 2020, Volume: 77 Issue: 1, 15 - 24, 01.03.2020

Aysegul Gozalan Özlem Ünaldı Fisun Kıırca , Nilay Çöplü Tuba Muderris Ziya Cibali Acikgoz Rıza Durmaz

Abstract

Amaç: Bu çalışmada; yoğun bakım ünitelerinde takip edilen hastaların kan örneklerinden izole edilen karbapenem dirençli Acinetobacter baumannii izolatları arasındaki klonal ilişki ve karbapenem direnç genlerinin moleküler yöntemler ile araştırılması amaçlandı. Yöntem: Kan kültürü şişelerinden bakteri izolasyonu için Bactec 9240 sistemi Becton Dickinson, ABD kullanıldı. Çalışmaya; konvansiyonel testler, API 20NE bioMèrieux, Fransa ve Phoenix TM 100 sistemi Becton Dickinson, ABD ile tanımlanan ve blaOXA-51 gen varlığı gösterilerek doğrulanan 112 A. baumannii suşu dahil edildi. Antimikrobiyal duyarlılık testleri Kirby-Bauer disk difüzyon yöntemi ve Phoenix TM 100 sistemi ile gerçekleştirildi. Karbapenem direnç genleri; blaOXA-23, blaOXA-48, blaOXA-58, blaIMP, blaVIM ve blaNDM-1 Multipleks Polimeraz Zincir Reaksiyonu PZR yöntemi ile araştırıldı. Acinetobacter baumannii suşları arasındaki klonal ilişkinin belirlenmesi için Pulse Field Jel Elektroforezi PFGE yöntemi kullanıldı. Bulgular: Suşların antibiyotik direnç yüzdeleri gentamisin, amikasin, tobramisin, netilmisin, seftazidim, trimetoprim/sulfametoksazol, piperasilin, siprofloksasin, ampisilin/sulbaktam, piperasilin/ tazobaktam ve sefoperazon/sulbaktam için sırasıyla %88; %81; %78; %36; %98,5; %96; %89; %100; %100; %93; %91 olarak bulundu. İmipenem ve meropenem MİK değerleri tüm grupta ≥8 µg/mL’nin üzerinde saptandı. Çalışmaya alınan izolatların tümünde blaOXA-51 ve blaOXA-23 gen varlığı tespit edildi. PFGE yöntemi ile 62 farklı pulsotip saptandı. Pulsotiplerden 19 tanesi birbiriyle ayırt edilemez profil gösteren eşittir üzeri ≥2 suş içermekteydi. Toplam 108 %96,4 suşun benzerlik oranı pulsotipler için %85 ve üzeri kabul edildiği durumda klonal yönden ilişkili 11 grup içerisinde toplandıkları gözlendi. Sonuç: Bu çalışmada, karbapenem dirençli A. baumannii suşlarının netilmisin dışında çalışılan tüm antibiyotiklere çok yüksek yüzdelerle dirence sahip olduğu ve hastane içinde çapraz bulaş yoluyla yayıldığı gösterildi. Bu suşlar hastane enfeksiyonları açısından risk oluşturmaktadır, bunula birlikte klonal yönden ilişkili suşlar spesifik bir ünite ve zaman periyodu ile sınırlı değildir.

Keywords

Acinetobacter baumannii, antibiyotik direnci, yoğun bakım üniteleri, polimeraz zincir reaksiyonu, pulsed-field jel elektroforezi

Molecular characterisation of Carbapenem-resistant Acinetobacter baumannii bloodstream infections in intensive care units

Abstract

Objective: In this study, the aim was to investigate the clonal relationship between carbapenem resistant Acinetobacter baumannii isolates and carbapenem resistance genes isolated from blood samples of patients followed in intensive care units by molecular methods. Methods: Bactec 9240 system Becton Dickinson, USA was used for the isolation of bacteria from blood culture flasks. Identification of 112 strains included in the study were performed by conventional tests, API 20NE bioMèrieux, France and Phoenix TM 100 system Becton Dickinson, USA and confirmed by the presence of blaOXA-51 gene. Antimicrobial susceptibility tests were performed by Kirby-Bauer disk diffusion method and Phoenix TM 100 system. Carbapenem resistance genes; blaOXA-23, blaOXA-48, blaOXA-58, blaIMP, blaVIM and blaNDM-1 were investigated by Multiplex Polymerase Chain Reaction PCR method. Pulsed Field Gel Electrophoresis PFGE was used to determine the clonal relationship between Acinetobacter baumannii strains. Results: The antibiotic resistance percentages of strains for gentamicin, amikacin, tobramycin, netil micin,ceftazidime,trimethoprim/sulfamethoxazole,piperacillin, ciprofloxacin, ampicillin/sulbactam, piperacillin/tazobactam and cefoperazone/ sulbactam, were 88%; 81%; 78%; 36%; 98.5%; 96%; 89%; 100%; 100%; 93%; 91% respectively. MIC values of imipenem and meropenem were determined above ≥8 µg/ml in the whole group. blaOXA-51 and blaOXA-23 genes were detected in all isolates included in the study. By PFGE method, 62 different pulsotypes were detected. Among the pulsotypes, 19 of them contained ≥2 strains. It was observed that 108 96.4% strains were clustered in 11 clonally related groups when the similarity between pulsotypes for grouping was limited to 85% or more. Conclusion: In this study, it was observed that carbapenem-resistant A. baumannii strains were resistant for all tested antibiotics at high levels except netilmicin and spread in the hospital via cross contamination. These strains posed a risk for hospital infections, however, clonal-related strains were not limited to a specific unit and time period.

Keywords

Acinetobacter baumannii, antibiotic resistance, Intensive care units, Polymerase Chain Reaction, pulsed-field gel electrophoresis

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