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

Yıl 2018, Cilt: 39 Sayı: 4, 1063 - 1068, 24.12.2018

Azer Özad Düzgün Ayşegül Saral

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

Cited By: 1

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-1 4 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

Öz

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.

Anahtar Kelimeler

beta-lactamase, Antibiotic resistance, integron

Kaynakça

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