Molecular Analysis of Resistance Genes in Multidrug-Resistant Klebsiella Pneumoniae İsolates

Abstract

Purpose: Development of antimicrobial resistance in Kelbsiella pneumoniae is a major problem in our country as well as in the whole world. Resistant K. pneumoniae is a bacterium that the World Health Organization has given critical priority in the list of the most dangerous pathogens for which antimicrobial drug development is needed. The aim of our study was to investigate carbapenem and colistin resistance genes in multidrug resistant K. pneumoniae isolates in a tertiary hospital in Turkey. Materials and Methods: Identification and antibiotic susceptibility tests were performed by Vitek-2 (bioMerieux, France). results for carbapenem and colistin were confirmed by gradient diffusion and microdilution methods. Carbapenemase and mcr genes were investigated by Polymerase Chain Reaction (PCR). Results: The blaNDM-1, blaOXA-48, blaKPC, blaVIM and blaIMP genes that cause resistance to carbapenem group antibiotics were tried to be determined by PCR in 69 isolates. As a result of the experiments, 54 (78%) of 69 isolates were positive for blaOXA-48 and 13 (19%) for blaNDM-1. KPC, VIM and IMP type beta-lactamases were not detected in any isolate. NDM-1 and OXA-48 type carbapenemases were found to be coexisting in 11 K. pneumoniae isolates. The mcr-1, mcr-2, mcr-3, mcr-4 and mcr-5 genes were not detected in any of the colistin resistant isolates. Conclusion: Colistin and carbapenem resistance mechanisms should be investigated by molecular methods in tertiary hospitals. The resistance profile data from the present study can be used as a knowledge base to understand the prevalent infection pattern in the hospital.

Keywords

• carbapenem
• carbapenemase
• colistin
• mcr

Supporting Institution

This study was supported by the Amasya University Scientific Research Projects (BAP) Unit under project number FMB-BAP-22-0547.

Project Number

FMB-BAP 22-0547

Ethical Statement

This study was approved by Amasya University Clinical Research Ethics Committee (Date: dated 07.07.2022 and numbered 79).

Çok İlaca Dirençli Klebsiella Pneumoniae İzolatlarında Direnç Genlerinin Moleküler Analizi

Abstract

Amaç: Klebsiella pneumoniae’da antimikrobiyal direnç gelişimi tüm dünyada olduğu gibi ülkemizde de büyük bir problemdir. Dirençli K. pneumoniae, Dünya Sağlık Örgütünün antimikrobiyal ilaç geliştirmeye ihtiyaç duyulan en tehlikeli patojenlerin listesinde kritik öncelik verdiği bir bakteridir. Bu çalışmanın amacı, Türkiye'de üçüncü basamak bir hastanede çok ilaca dirençli K. pneumoniae izolatlarında karbapenem ve colistin direnç genlerinin araştırılmasıdır. Araçlar ve Yöntem: Tanımlama ve antibiyotik duyarlılık testleri Vitek-2 (bioMerieux, Fransa) tarafından gerçekleştirildi. Karbapenem ve kolistin için sonuçlar gradyan difüzyon ve mikrodilüsyon yöntemleriyle doğrulandı. Karbapenemaz ve mcr genleri Polimeraz Zincir Reaksiyonu (PCR) ile araştırıldı. Karbapenem grubu antibiyotiklere direnç oluşumunu sağlayan blaNDM-1, blaOXA-48, blaKPC, blaVIM ve blaIMP genleri 69 izolatta çalışıldı. Bulgular: Yapılan deneyler sonucunda 69 izolatın 54’ünde (%78) blaOXA-48 ve 13’ünde (%19) blaNDM-1 pozitif olduğu tespit edildi. Hiçbir izolatta KPC, VIM ve IMP tipi beta-laktamaz belirlenemedi. Çalışmamızda 11 (%16) K. pneumoniae izolatında NDM-1 ve OXA-48 tipi karbapenemazın birlikte bulunduğu saptandı. Kolistin dirençli izolatların hiçbirinde mcr-1, mcr-2, mcr-3, mcr-4 ve mcr-5 genleri tespit edilememiştir. Sonuç: Üçüncü basamak hastanelerde kolistin ve karbapenem direnç mekanizmalarını moleküler yöntemlerle araştırılmalıdır. Mevcut çalışmadan elde edilen direnç profili verileri, hastanede yaygın olan enfeksiyon örüntüsünün anlaşılması için bir bilgi tabanı olarak kullanılabilir.

Keywords

• karbapenem
• karbapenemaz
• kolistin
• mcr

Supporting Institution

Bu çalışma, Amasya Üniversitesi Bilimsel Araştırma Projeleri (BAP) Birimi tarafından FMB-BAP-22-0547 numaralı proje kapsamında desteklenmiştir.

Project Number

FMB-BAP 22-0547

Ethical Statement

Bu çalışma Amasya Üniversitesi Klinik Araştırma Etik Kurulu tarafından onaylanmıştır (Tarih: 07.07.2022, No: 79).

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