Çoklu-İlaç Dirençli P. aeruginosa ve E.coli Suşlarında Antibiyotik Direnç Genlerinin ve Sınıf 1 İntegron Gen Kasetlerinin Araştırılması

Azer Özad Düzgün; Ayşegül Saral

doi:10.17776/csj.409136

TR EN

Çoklu-İlaç Dirençli P. aeruginosa ve E.coli Suşlarında Antibiyotik Direnç Genlerinin ve Sınıf 1 İntegron Gen Kasetlerinin Araştırılması

Öz

P. aeruginosa ve E.coli izolatlarının birçok antibiyotiğe direnç kazanması nedeniyle bu patojenlerin neden olduğu enfeksiyonların tedavisi zorlaşmıştır. Bundan dolayı klinik izolatlarda antibiyotik direnç genlerinin belirlenmesi büyük önem arz etmektedir. Bu çalışmanın amacı çoklu-ilaç dirençli P. aeruginosa ve E.coli klinik suşlarında beta laktamaz direnç genlerinin ve sınıf 1 integronlar gen kasetlerinin varlığının araştırılmasıdır. Gümüşhane Devlet Hastanesi yoğun bakım ünitesinde yatan hastaların kan, idrar ve balgam örneklerinden izole edilen 2 P. aeruginosa ve 2 E.coli izolatlarının tanımlaması ve antibiyogramı Vitek 2 Compact otomatize sistemi ile çalışılmıştır. Total DNA izolasyonu kaynatma DNA metoduyla yapılmıştır. bla_VIM, bla_NDM, bla_IMP, bla_GES, bla_CTXM-1, bla_CTXM-2, bla_OXA-58, bla_OXA-23, bla_OXA-51, bla_OXA-40, bla_KPC ve sınıf 1 integron gen kasetlerine ait primerler kullanılarak PZR’ler gerçekleştirilmiştir. Amplikasyonun gerçekleştiği tüm örnekler %1’lik agaroz jelde yürütülmüştür ve daha sonra UV ışığında görünür hale getirilmiştir. Antibiyogram sonucuna göre P. aeruginosa izolatları gentamisin hariç kullanılan tüm antibiyotiklere karşı direnç gösterdiği belirlenmiştir. E.coli izolatlarının ise karbapenemlere karşı duyarlı sefalosporin grubu antibiyotiklere karşı dirençli olduğu görülmüştür. Sınıf A beta laktamaz genlerinden bla_CTXM-14 suşun 3’ünde bulunurken, hiçbir suşta bla_CTXM-2, bla_KPC ve bla_GES genleri tespit edilmemiştir. Araştırılan sınıf B ve sınıf D beta laktamaz genlerinin varlığı hiçbir örnekte görülmemiştir. 3 suşta sınıf 1 integronun varlığı tespit edilmiştir. DNA dizi analiz sonucuna göre üç integron pozitif örneğin dfrA17/AadA5 gen kasetine sahip olduğu belirlenmiştir.

Anahtar Kelimeler

beta-laktamaz,Antibiyotik direnci,integron

Investigation of Antibiotic Resistance Genes and Class 1 Integron in Multi-Drug Resistant P. aeruginosa and E. coli Strains

Abstract

P. aeruginosa and E. coli are resistant to many antibiotics, so the treatment of infections of these pathogens has become difficult. Therefore, searching for the presence of antibiotic resistance genes in clinical isolates is of great importance. The purpose of this study is to investigate the presence of beta-lactam resistance genes and class 1 integron in multi-drug resistant P. aeruginosa and E. coli clinical strains. Vitek 2 Compact automatization system were used for identification and antibiogram of 2 P. aeruginosa and 2 E. coli isolates which were isolated from blood, urine and sputum specimens of patients whom were hospitalized in Gümüşhane State Hospital intensive care unit. Total DNA isolation was done by boiling DNA method. PCR were performed using primers of class 1 integron gene cassette, bla_VIM, bla_NDM,bla_IMP, bla_GES, bla_CTXM-1, bla_CTXM-2, bla_OXA-58, bla_OXA-23, bla_OXA-51, bla_OXA-40 and bla_KPC. All amplification samples were performed on 1% agarose gel and subsequently visualized under UV light. According to the results of the antibiogram, P. aeruginosa isolates showed resistance against all used antibiotics except gentamicin. The E. coli isolates were found to be resistant to cephalosporin antibiotics and susceptible to carbapenems. bla_CTXM-1-class A beta lactamase genes was found in 3 strains of 4, while no bla_CTXM-2,bla_GES and bla_KPC genes were detected in the strains. The presence of the investigated class B and class D beta lactamase genes was not observed in any sample. The presence of class 1 integron was detected in 3 strains. According to the result of the DNA sequence analysis, it was determined that three integron positive samples had the dfrA1/AadA5 gene cassettes.

Keywords

beta-lactamase,Antibiotic resistance,integron

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Details

Primary Language

Turkish

Subjects

-

Journal Section

Research Article

Authors

Azer Özad Düzgün ^*
Türkiye

Ayşegül Saral

Publication Date

December 24, 2018

Submission Date

March 23, 2018

Acceptance Date

October 19, 2018

Published in Issue

Year 1970 Volume: 39 Number: 4

DOI

https://doi.org/10.17776/csj.409136

IZ

https://izlik.org/JA34HG59FZ

Cite

RIS / Bibtex

APA

Özad Düzgün, A., & Saral, A. (2018). Çoklu-İlaç Dirençli P. aeruginosa ve E.coli Suşlarında Antibiyotik Direnç Genlerinin ve Sınıf 1 İntegron Gen Kasetlerinin Araştırılması. Cumhuriyet Science Journal, 39(4), 1063-1068. https://doi.org/10.17776/csj.409136

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